Information on EC 3.4.21.105 - rhomboid protease and Organism(s) Escherichia coli and UniProt Accession P09391

commercially available fluorescent substrate

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?

Gurken protein + H2O

?

-

-

?

protein Bla-LY2-MBP + H2O

?

recombinantly expressed type I model membrane protein substrate having the second transmembrane region of lactose permease LY2 at the extramembrane region in vivo and in vitro at the predicted periplasm-membrane boundary region of LY2, the determinants for proteolysis reside within the LY2 sequence, GlpG cleaves an extramembrane region of the substrate exposed to the periplasm, overview

683901

-

?

protein MIC2 + H2O

?

cleavage at an Ala-Gly bond

683916

-

?

reporter substrate LY2

?

using a combinatorial approach it is shown that a negatively charged residue is the primary determinant of cleavage. The amino acid preceding peptide bond hydrolysis (the P1 position) has a preference for the small and polar Ser residue. The amino acid succeeding peptide bond hydrolysis (the P1 position) has a preference for negatively charged Asp

710566

-

?

Spitz-polyA + H2O

?

-

-

?

TatA + H2O

?

-

-

?

TatA protein + H2O

?

-

-

?

3,4-dichloroisocoumarin + H2O

?

a significant portion of the inhibitor 3,4-dichloroisocoumarin bound to GlpG is enzymatically turned over

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?

beta-lactamase Spitz transmembrane domain + H2O

?

-

34 residue peptide, sequence KRPRPMLEKASIASGAMCALVFMLFVCLAFYLRK

732381

-

?

BODIPY FL casein + H2O

?

-

-

?

C100Spi-Flag + H2O

?

-

no cleavage of C100-Flag

-

?

FL-casein + H2O

?

-

-

?

Gurken + H2O

?

-

-

?

Gurken protein + H2O

PQRKVRMA + HIVFSFFV

-

-

?

Keren + H2O

?

-

-

?

LacYTM2 protein + H2O

DINHISKS + DTGIIFAA

-

-

?

N-acetyl-PEG4-QRKVRMAHIVFSFPC-amide + H2O

N-acetyl-PEG4-QRKVRMA + HIVFSFPC-amide

-

i.e. peptide KSp21

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?

Protein + H2O

?

-

cleaves a model protein having an N-terminal and periplasmically localized beta-lactamase domain, a LacY-derived transmembrane region, and a cytosolic maltose binding protein mature domain, cleavage occurs between Ser and Asp in a region of high local hydrophilicity, which might be located iin a juxtamembrane rather than an intramembrane position. The conserved Ser and His residue of GlpG are esential for proteolytic activity

658102

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?

Spitz + H2O

?

-

-

?

TatA + H2O

?

-

transmembrane substrate from Providencia stuartii. Binding of TatA occurs with positive cooperativity in an exosite-mediated mode of substrate binding. Exosite formation is dependent on the oligomeric state of rhomboids, and when dimers are dissociated, allosteric substrate activation is not observed

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?

TatA protein + H2O

MESTIATA + AFGSPWQL

-

-

?

additional information

?

-

18 entries

Gurken protein + H2O

?

-

-

?

TatA protein + H2O

?

-

-

?

Gurken protein + H2O

PQRKVRMA + HIVFSFFV

-

-

?

LacYTM2 protein + H2O

DINHISKS + DTGIIFAA

-

-

?

TatA protein + H2O

MESTIATA + AFGSPWQL

-

-

?

additional information

?

-

5 entries

additional information

-

enzyme does not require cofactors to catalyze intramembrane proteolysis

-

2-methylpropyl 2-oxo-4-phenylazetidine-1-carboxylate

beta-lactam inhibitor, forms a single bond to the catalytic serine and the carbonyl oxygen of the inhibitor faces away from the oxyanion hole. The hydrophobic N-substituent of the inhibitor points into a cavity within the enzyme, providing a structural explanation for the specificity of beta-lactams on rhomboid proteases. This same cavity probably represents the S2' substrate binding site

3,4-dichloroisocoumarin

2 entries

3-butyl-4-(pent-4-yn-1-yl)oxetan-2-one

-

4-chloro-7-nitro-3-[(5-phenylpentyl)oxy]-1H-2-benzopyran-1-one

inhibitor reacts with virtually all tested rhomboids

7-amino-3-butoxy-4-chloro-1H-isochromen-1-one

-

7-amino-4-chloro-3-(2-phenylethoxy)-1H-isochromen-1-one

-

7-amino-4-chloro-3-methoxyisocoumarin

-

7-amino-4-chloro-3-[(5-phenylpentyl)oxy]-1H-isochromen-1-one

-

cyclopentyl 2-oxo-4-phenylazetidine-1-carboxylate

beta-lactam inhibitor, forms a single bond to the catalytic serine and the carbonyl oxygen of the inhibitor faces away from the oxyanion hole. The hydrophobic N-substituent of the inhibitor points into a cavity within the enzyme, providing a structural explanation for the specificity of beta-lactams on rhomboid proteases. This same cavity probably represents the S2' substrate binding site

diisopropyl fluorophosphonate

mechansim-based inhibitor

phenyl 2-oxo-4-phenylazetidine-1-carboxylate

beta-lactam inhibitor, forms a single bond to the catalytic serine and the carbonyl oxygen of the inhibitor faces away from the oxyanion hole. The hydrophobic N-substituent of the inhibitor points into a cavity within the enzyme, providing a structural explanation for the specificity of beta-lactams on rhomboid proteases. This same cavity probably represents the S2' substrate binding site

(3S,4S)-1-[(4-chlorophenyl)sulfonyl]-3-methyl-4-phenylazetidin-2-one

-

(3S,4S)-3-butyl-4-(pent-4-yn-1-yl)oxetan-2-one

-

753260

1-(2,3-dihydro-4H-1,4-benzoxazin-4-yl)-3,3,3-trifluoro-2-(trifluoromethyl)propan-1-one

-

1-(biphenyl-3-ylsulfonyl)-4-phenylazetidin-2-one

-

1-(biphenyl-4-ylsulfonyl)-4-phenylazetidin-2-one

-

1-[(3'-methylbiphenyl-4-yl)sulfonyl]-4-phenylazetidin-2-one

-

1-[(3-bromophenyl)sulfonyl]-4-phenylazetidin-2-one

-

1-[(3-chlorophenyl)sulfonyl]-4-(2-phenylethyl)azetidin-2-one

-

1-[(3-chlorophenyl)sulfonyl]-4-(propan-2-yl)azetidin-2-one

-

1-[(4'-chlorobiphenyl-4-yl)sulfonyl]-4-phenylazetidin-2-one

-

1-[(4-bromophenyl)sulfonyl]-4-phenylazetidin-2-one

-

1-[(4-chlorophenyl)sulfonyl]-3-methylazetidin-2-one

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1-[(4-methylphenyl)sulfonyl]-4-phenylazetidin-2-one

-

2-(benzyloxy)-5-chloro-4H-3,1-benzoxazin-4-one

-

covalent, but slow reversible inhibition mechanism

753003

2-(benzyloxy)-5-methyl-4H-3,1-benzoxazin-4-one

-

covalent, but slow reversible inhibition mechanism

753003

3,3,3-trifluoro-N-[(5-methyl-2-phenyl-2H-1,2,3-triazol-4-yl)methyl]-2-(trifluoromethyl)propanamide

-

3,3,3-trifluoro-N-[2-(propan-2-yloxy)phenyl]-2-(trifluoromethyl)propanamide

-

3,4-dichloro-1H-2-benzopyran-1-one

-

753260

3,4-dichloroisocoumarin

-

3-methyl-1-[(4-methylphenyl)sulfonyl]-4-phenylazetidin-2-one

-

4-(2-chlorophenyl)-1-[(3-chlorophenyl)sulfonyl]azetidin-2-one

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4-(3-bromophenyl)-1-[(3-chlorophenyl)sulfonyl]azetidin-2-one

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4-[(3-methyl-2-oxoazetidin-1-yl)sulfonyl]benzonitrile

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acetyl-L-Ile-L-Ala-L-Thr-L-Ala-chloromethylketone

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inhibitor derived from the natural rhomboid substrate TatA from bacterium Providencia stuartii, binds in a substrate-like manner

acetyl-L-Phe-L-Ala-L-Thr-L-Ala-chloromethylketone

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inhibitor derived from the natural rhomboid substrate TatA from bacterium Providencia stuartii, binds in a substrate-like manner

benzyl (2S)-1-[(4-methylphenyl)sulfonyl]-4-oxoazetidine-2-carboxylate

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dichloroisocoumarin

-

below 0.1 mM

diisopropyl fluorophosphonate

irreversible inhibition

N-(2,6-dimethylphenyl)-3,3,3-trifluoro-2-(trifluoromethyl)propanamide

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N-[2-(cyclopentyloxy)phenyl]-3,3,3-trifluoro-2-(trifluoromethyl)propanamide

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N-[2-(cyclopropylmethoxy)phenyl]-3,3,3-trifluoro-2-(trifluoromethyl)propanamide

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phenyl 2-oxo-4-phenylazetidine-1-carboxylate

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tert-butyl 2-[[3,3,3-trifluoro-2-(trifluoromethyl)propanoyl]amino]benzoate

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additional information

8 entries

-

0.00074 - 0.0029

BODIPY FL casein

12 entries

-

0.0025

FL-casein

-

pH 6.0, 3°C

-

0.022

TatA

-

K0.5 value, Hill coefficient 1.2, pH 6.0, 3°C

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0.003 - 0.0073

BODIPY FL casein

12 entries

-

0.062

FL-casein

-

pH 6.0, 3°C

-

0.0017

TatA

-

pH 6.0, 3°C

-

2.4 - 7.78

BODIPY FL casein

12 entries

-

2.48

FL-casein

-

pH 6.0, 3°C

-

0.08

TatA

-

pH 6.0, 3°C

-

0.044

3-butyl-4-(pent-4-yn-1-yl)oxetan-2-one

Escherichia coli

pH not specified in the publication, temperature not specified in the publication

0.0004

7-amino-3-butoxy-4-chloro-1H-isochromen-1-one

Escherichia coli

pH 7.3, 37°C

0.0011

7-amino-4-chloro-3-(2-phenylethoxy)-1H-isochromen-1-one

Escherichia coli

pH 7.3, 37°C

0.006

7-amino-4-chloro-3-methoxyisocoumarin

Escherichia coli

in 50 mM HEPES-NaOH (pH 7.5), 0.4 M NaCl, 5 mM EDTA, 10% (v/v) glycerol, and 0.05% (w/v) n-dodecyl-beta-D-maltoside, at 37°C

0.00075

7-amino-4-chloro-3-[(5-phenylpentyl)oxy]-1H-isochromen-1-one

Escherichia coli

pH 7.3, 37°C

0.029 - 0.044

(3S,4S)-3-butyl-4-(pent-4-yn-1-yl)oxetan-2-one

2 entries

0.001 - 0.0012

2-(benzyloxy)-5-methyl-4H-3,1-benzoxazin-4-one

2 entries

0.0056 - 0.019

3,4-dichloro-1H-2-benzopyran-1-one

5.7 - 6.5

-

25 - 37

-

much more active az 37°C than at 25°C

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-

membrane-embedded, traverses the membrane six times

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658102

membrane

3 entries

physiological function

studies in bacteria and Drosophila show that RHO subfamily members Aar from Providencia stuarti and Rho-1 from Drosophila melanogaster can functionally complement each other

710566

physiological function

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rhomboid displays a rapid diffusion comparable to the diffusion of small, single-pass transmembrane proteins, with a diffusion coefficient of about 0.8 microm2 per s. The rhomboid fold is able to distort surrounding lipids, overcome the viscosity limit of the membrane, and accelerate its search for substrates

141500

-

protein with detergent bound, sedimentation equilibrium result

31772

-

2 * 31772, native protein, 2 * 71615, protein with detergent bound, calculated

71615

-

2 * 31772, native protein, 2 * 71615, protein with detergent bound, calculated

dimer

-

2 * 31772, native protein, 2 * 71615, protein with detergent bound, calculated

additional information

10 entries

analysis of interaction energies among the active site residues His254, Ser201, and Asn154, which form a hydrogen bonding network. In mild detergent, the active site residues are weakly coupled with interaction energies of ?1.4 kcal/mol between His254 and Ser201 and ?0.2 kcal/mol between Ser201 and Asn154. These residues are important for function and also for the folding cooperativity of GlpG. The weak interaction between Ser and His in the catalytic dyad may partly explain the unusually slow proteolysis by GlpG

crystal structure in complex with beta-lactam inhibitors phenyl 2-oxo-4-phenylazetidine-1-carboxylate, 2-methylpropyl 2-oxo-4-phenylazetidine-1-carboxylate, and cyclopentyl 2-oxo-4-phenylazetidine-1-carboxylate, to 2.2 to 2.4 A resolution

crystal structure of the soluble cytoplasmic domain, 1.35 Å resolution. The cytoplasmic domain exists as a dimer with extensive domain swapping between the two monomers. Domain-swapped dimers can be isolated from the full-length protein

crystals of the truncated GlpG wild type enzyme are obtained by mixing 2.5-3 M ammonium chloride with protein at ratio of 1:1 in hanging drops at 25°C

GlpG in a more open conformation, where the capping loop L5 has been lifted, exposing the previously buried and catalytically essential Ser201 to outside aqueous solution, X-ray diffraction structure determination and analysis at 2.5-2.6 A resolution

684010

in complex with covalently bound inhibitor 3-butyl-4-(pent-4-yn-1-yl)oxetan-2-one

in complex with inhibitors 3,4-dichloroisocoumarin and diisopropyl fluorophosphonate, to 2.3 A resolution. Enzyme forms a covalent adduct with diisopropyl fluorophosphonate, which mimics the oxyanion-containing tetrahedral intermediate of the hydrolytic reaction. The oxyanion is stabilized by the main chain amide of residue Ser201 and by the side chains of His150 and Asn154. The phosphorylation of the catalytic Ser201 weakens its interaction with His254, causing the catalytic histidine to rotate away from the serine and accompanied by further rearrangement of the side chains of Tyr205 and Trp236 within the substrate-binding groove and by opening of the L5 cap and movement of transmembrane helix S5 toward S6 in a direction different from that predicted by the lateral gating model

molecular dynamics simulation of substrate entry into the active site. Substrate is inclined to enter into the active site along a path between Loop3 and Loop5, and residue His150 plays an important role in substrate entry

754206

molecular dynamics simulation. In both membrane and lipid-solubilized environments the S201/H254 and S201/H150 interatomic distances of GlpG are most sensitive to variations of the protonation state of the active site residues. The catalytic diad of the lipid-solubilized enzyme exists as an H254(+)-S201(-) ion pair at the Michaelis complex stage, with Ser201 ready for nucleophilic attack on the substrate. Therefore, deprotonation of S201 does not contribute to the activation barrier of covalent tetrahedral complex formation. Both catalytic residues, H254 and S201, are neutral in the Michaelis complex of GlpG in the membrane. Therefore, S201 deprotonation by H254 general base catalysis should contribute to the activation barrier of the covalent tetrahedral complex formation

732230

molecular dynamics simulations initiated from both gate-open and gate-closed states of rhomboid GlpG in a phospholipid bilayer. There is rapid loss of crystallographic waters from the active site, but retention of a water cluster within a site formed by His141, Ser181, Ser185, and/or Gln189. Residues Gln189 and Ser185 play an essential role in catalysis with no effect on structural stability

purified GlpG core domain, hanging drop vapour diffusion method, room temperature, 5 mg/ml membrane protein in 10 mM Tris-HCl, pH 7.6, and 20 mM nonylglucoside, over a reservoir solution of 3 M NaCl and 100 mM Bis-Tris propane, pH 7.0, cryoprotection by 25% glycerol, 1 month, X-ray diffraction structure determination and analysis at 2.1 A resolution

683916

purified recombinant His-tagged GlpG, hanging drop vapour diffusion method, 5 mg/ml protein in 20 mM HEPES, pH 7.5, 90 mM NaCl, 10% glycerol, and lauryl dimethylamine oxide, mixed with a reservoir solution containing 30% w/v PEG 400, 200 mM CaCl2, and 100 mM MES, pH 6.5, X-ray diffraction structure determination and analysis at 2.25-2.3 A resolution

684012

structure of Escherichia coli GlpG consists of a 6-transmembrane domain topology

710565

Coarse-grained molecular dynamics simulations in hydrated lipid bilayers to study the interaction of rhomboid protease GlpG with the transmembrane domain of the substrate Spitz. Spitz does not associate with GlpG exclusively at the putative substrate gate near TMD 5. Instead, there are six prominent and stable interaction sites, including one between TMDs 1 and 3, with the closest enzyme-substrate proximity occurring at the ends of helical transmembrane domains or in loops. The initial interaction between enzyme and substrate is not limited to a single site on the enzyme, and may be driven by juxtamembrane electrostatic interactions

-

732381

in complex with inhibotrs acetyl-L-Ile-L-Ala-L-Thr-L-Ala-chloromethylketone, acetyl-L-Phe-L-Ala-L-Thr-L-Ala-chloromethylketone. Inhibitors bind in a substrate-like manner. The S1 subsite is prominent and merges into the water retention site, suggesting intimate interplay between substrate binding, specificity and catalysis. The S4 subsite is plastically formed by residues of the L1 loop, an important but hitherto enigmatic feature of the rhomboid fold

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rhomboid protease GlpG in complex with 3,4-dichloroisocoumarin and diisopropyl fluorophosphonate, sitting drop vapor diffusion method, using 100 mM Bis-Tris propane (pH 7.0), and 3 M NaCl

A253I

the mutant exhibits 16% of wild type activity towards the wild type TatA protein but shows 87% of wild type activity on TatA mutant A8G

A253L

the mutant exhibits no activity of wild type activity towards the wild type TatA protein but shows 23% of wild type activity on TatA mutant A8G

A253T

the mutant exhibits 37% of wild type activity towards the wild type TatA protein but shows 63% of wild type activity on TatA mutant A8G

A253V

the mutant exhibits 63% of wild type activity towards the wild type TatA protein but shows 144% of wild type activity on TatA mutant

D18A

mutation in residue conserved among 32 sequenced prokaryotic rhomboids. No significant change in activity is observed

D243A

site-directed mutagenesis, the mutant shows similar activity as the wild-type enzyme

E42A

mutation in residue conserved among 32 sequenced prokaryotic rhomboids. No significant change in activity is observed

F133Y/F135Y

site-directed mutagenesis, almost inactive mutant

F139S

site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme

F153A/W236A

site-directed mutagenesis, the enzyme shows 10fold increased activity compared to the wild-type enzyme

F245A

site-directed mutagenesis, the enzyme shows increased activity compared to the wild-type enzyme

G199A

site-directed mutagenesis, inactive mutant

G257A

site-directed mutagenesis, inactive mutant

H141F

decrease in transition temperature by 6 degrees. Mutant retains almost no activity

H141T

decrease in transition temperature by 11-12 degrees. Mutant retains some activity

H141V

decrease in transition temperature by 11-12 degrees. Mutant retains some activity

H145A

site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme

H150A

the mutation leads to a complete loss of activity

H2541X

using mutagenesis it is shown that His254 is catalytically essential

H254A

L143S

site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme

L229V/F232V/W236V

L244A

site-directed mutagenesis, the mutant shows similar activity as the wild-type enzyme

M247A

site-directed mutagenesis, the mutant shows similar activity as the wild-type enzyme

M249A

site-directed mutagenesis, the enzyme shows increased activity compared to the wild-type enzyme

M3A

mutation in residue conserved among 32 sequenced prokaryotic rhomboids. No significant change in activity is observed

N154A

N154A/H254A

mutation induces larger destabilization

N154A/S201A

mutation induces larger destabilization

N251A

site-directed mutagenesis, inactive mutant

N33P

mutation promotes domain-swapped dimer formation, due to probably a lower entropic barrier of proteinprotein association

Q14A

mutation in residue conserved among 32 sequenced prokaryotic rhomboids. No significant change in activity is observed

Q189A

no catalytic activity, thermostability of mutant is indistinguishable from wild-type

Q189T

no catalytic activity, thermostability of mutant is indistinguishable from wild-type

Q30A

mutation in residue conserved among 32 sequenced prokaryotic rhomboids. No significant change in activity is observed

R11A

mutation in residue conserved among 32 sequenced prokaryotic rhomboids. No significant change in activity is observed

R137A

site-directed mutagenesis, inactive mutant

R49A

mutation in residue conserved among 32 sequenced prokaryotic rhomboids. No significant change in activity is observed

S185T

mutant retains proteolyitc activity

S185V

transition temperature similar to wild-type, no catalytic activity

S201A

S201A/H254A

double mutation on the catalytic dyad, yields a smaller decrease in the stability than individual single mutations

S201X

using mutagenesis it is shown that Ser201 is catalytically essential

710565

S68A

mutation in residue conserved among 32 sequenced prokaryotic rhomboids. No significant change in activity is observed

T22A

mutation in residue conserved among 32 sequenced prokaryotic rhomboids. No significant change in activity is observed

W136A

site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme

W157A/F232A

site-directed mutagenesis, the enzyme shows 6fold increased activity compared to the wild-type enzyme

W157C/F232C

site-directed mutagenesis, the enzyme shows reduced activity compared to the wild-type enzyme

W38A

mutation in residue conserved among 32 sequenced prokaryotic rhomboids. No significant change in activity is observed

Y138D

site-directed mutagenesis, inactive mutant

Y138F

site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme

Y138S

site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme

Y138S/F139S

site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme

Y138S/F139S/L143S

Y138Y

site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme

Y160C/L229C

site-directed mutagenesis, the enzyme shows highly reduced activity compared to the wild-type enzyme

Y205A

site-directed mutagenesis, inactive mutant

S150A

-

inactive

S201A

-

inactive

additional information

4 entries

Ni-NTA column chromatography, Mono-Q column chromatography, and Superdex-200 gel filtration

recombinant His-tagged GlpG from membranes of strain C43(DE3) by nickel affinity chromatography

684012

use of polymers composed of maleic acid and either diisobutylene or styrene for solubilization of rhomboid proteases in lipid nanodiscs, with up to 48% efficiency. The activity of rhomboids in lipid nanodiscs is closer to that in the native membrane than rhomboids in detergent

-

His-Pure Cobalt affinity column chromatography

-

purification of the catalytic transmembrane domain of GlpG into phosphocholine or maltoside detergent micelles of varying alkyl chain lengths. Catalytic turnover numbers depend on detergent alkyl chain length, with saturated chains containing 10-12 carbon atoms supporting maximal activity. Similar results are obtained in phospholipid bicelles, with no proteolytic activity being detected in longer-chain lipids. Hydrophobic mismatch gives rise to a small change in structure and can exert an inhibitory effect on rhomboid activity

-

732886

-

4 entries

DNA and amino acid sequence analysis, phylogenetic analysis, overview

683501

expressed in Escherichia coli BL21 C43(DE3) cells

expression in Escherichia coli

expression of His-tagged GlpG in Escherichia coli strain C43(DE3)

-

-

expressed in Escherichia coli C43(DE3) cells

-

expression in Escherichia coli

-

731712, 753260

reconstitution of rhomboids in liposomes. Active rhomboids are reconstituted in large and giant unilamellar vesicles

-

analysis

screening assay using fluorescence polarization activity-based protein profiling

molecular biology

TM5 helix and L1 loop are dynamically coupled so that changes in the dynamics of one are relayed to the other

701252

658102

Maegawa, S.; Ito, K.; Akiyama, Y.

Proteolytic action of GlpG, a rhomboid protease in the Escherichia coli cytoplasmic membrane

Biochemistry

44

13543-13552

2005

Escherichia coli

16216077

Urban, S.; Schlieper, D.; Freeman, M.

Conservation of intramembrane proteolytic activity and substrate specificity in prokaryotic and eukaryotic rhomboids

Curr. Biol.

12

1507-1512

2002

Escherichia coli, Pseudomonas aeruginosa, Thermotoga maritima, Bacillus subtilis (P54493)

12225666

658553

Lemberg, M.K.; Menendez, J.; Misik, A.; Garcia, M.; Koth, C.M.; Freeman, M.

Mechanism of intramembrane proteolysis investigated with purified rhomboid proteases

EMBO J.

24

464-472

2005

Aquifex aeolicus, Bacillus subtilis, Drosophila sp. (in: flies), Escherichia coli, Providencia stuartii, Pseudomonas aeruginosa, Homo sapiens (Q9NX52)

15616571

Urban, S.; Wolfe, M.S.

Reconstitution of intramembrane proteolysis in vitro reveals that pure rhomboid is sufficient for catalysis and specificity

Proc. Natl. Acad. Sci. USA

102

1883-1888

2005

Aquifex aeolicus, Bacillus subtilis, Escherichia coli, Providencia stuartii

15684070

683499

Urban, S.

Rhomboid proteins: conserved membrane proteases with divergent biological functions

Genes Dev.

20

3054-3068

2006

Arabidopsis thaliana, Bacillus subtilis, Saccharomyces cerevisiae, Drosophila melanogaster, Escherichia coli, Homo sapiens, Providencia stuartii, Toxoplasma gondii

17114579

683501

Lemberg, M.K.; Freeman, M.

Functional and evolutionary implications of enhanced genomic analysis of rhomboid intramembrane proteases

Genome Res.

17

1634-1646

2007

Saccharomyces cerevisiae, Drosophila melanogaster, Homo sapiens, Mus musculus, Plasmodium falciparum, Toxoplasma gondii, Escherichia coli (P09391), Providencia stuartii (P46116), Bacillus subtilis (P54493), Pseudomonas aeruginosa (Q9HZC2)

17938163

683867

Lemberg, M.K.; Freeman, M.

Cutting proteins within lipid bilayers: rhomboid structure and mechanism

Mol. Cell

28

930-940

2007

Drosophila melanogaster, Escherichia coli, Haemophilus influenzae (P44783)

18158892

683901

Maegawa, S.; Koide, K.; Ito, K.; Akiyama, Y.

The intramembrane active site of GlpG, an E. coli rhomboid protease, is accessible to water and hydrolyses an extramembrane peptide bond of substrates

Mol. Microbiol.

64

435-447

2007

Escherichia coli (P09391), Escherichia coli

17493126

683913

Wu, Z.; Yan, N.; Feng, L.; Oberstein, A.; Yan, H.; Baker, R.P.; Gu, L.; Jeffrey, P.D.; Urban, S.; Shi, Y.

Structural analysis of a rhomboid family intramembrane protease reveals a gating mechanism for substrate entry

Nat. Struct. Mol. Biol.

13

1084-1091

2006

Escherichia coli

17099694

683916

Wang, Y.; Zhang, Y.; Ha, Y.

Crystal structure of a rhomboid family intramembrane protease

Nature

444

179-180

2006

Escherichia coli (P09391), Escherichia coli

17051161

684010

Wang, Y.; Ha, Y.

Open-cap conformation of intramembrane protease GlpG

Proc. Natl. Acad. Sci. USA

104

2098-2102

2007

Escherichia coli (P09391), Escherichia coli

17277078

684012

Ben-Shem, A.; Fass, D.; Bibi, E.

Structural basis for intramembrane proteolysis by rhomboid serine proteases

Proc. Natl. Acad. Sci. USA

104

462-466

2007

Escherichia coli (P09391), Escherichia coli

17190827

Baker, R.P.; Young, K.; Feng, L.; Shi, Y.; Urban, S.

Enzymatic analysis of a rhomboid intramembrane protease implicates transmembrane helix 5 as the lateral substrate gate

Proc. Natl. Acad. Sci. USA

104

8257-8262

2007

Escherichia coli (P09391)

17463085

701252

Bondar, A.N.; del Val, C.; White, S.H.

Rhomboid protease dynamics and lipid interactions

Structure

17

395-405

2009

Escherichia coli (P09391), Escherichia coli

19278654

710565

Ha, Y.

Structure and mechanism of intramembrane protease

Semin. Cell Dev. Biol.

20

240-250

2009

Saccharomyces cerevisiae, Drosophila melanogaster, Plasmodium falciparum, Toxoplasma gondii, Escherichia coli (P09391)

19059492

710566

Hill, R.B.; Pellegrini, L.

The PARL family of mitochondrial rhomboid proteases

Semin. Cell Dev. Biol.

21

582-592

2010

Danio rerio, Saccharomyces cerevisiae, Drosophila melanogaster, Homo sapiens, Mus musculus, Providencia stuartii, Schizosaccharomyces pombe, Escherichia coli (P09391)

20045481

Pierrat, O.A.; Strisovsky, K.; Christova, Y.; Large, J.; Ansell, K.; Bouloc, N.; Smiljanic, E.; Freeman, M.

Monocyclic beta-lactams are selective, mechanism-based inhibitors of rhomboid intramembrane proteases

ACS Chem. Biol.

6

325-335

2011

Escherichia coli, Providencia stuartii, Escherichia coli NR698

21175222

Vinothkumar, K.R.; Strisovsky, K.; Andreeva, A.; Christova, Y.; Verhelst, S.; Freeman, M.

The structural basis for catalysis and substrate specificity of a rhomboid protease

EMBO J.

29

3797-3809

2010

Escherichia coli (P09391), Escherichia coli

20890268

Xue, Y.; Ha, Y.

The catalytic mechanism of rhomboid protease GlpG probed by 3,4-dichloroisocoumarin and diisopropyl fluorophosphonate

J. Biol. Chem.

287

3099-3107

2012

Escherichia coli, Escherichia coli (P09391)

22130671

731319

Ghasriani, H.; Kwok, J.K.; Sherratt, A.R.; Foo, A.C.; Qureshi, T.; Goto, N.K.

Micelle-catalyzed domain swapping in the GlpG rhomboid protease cytoplasmic domain

Biochemistry

53

5907-5915

2014

Escherichia coli (P09391), Escherichia coli

25162988

Sampathkumar, P.; Mak, M.W.; Fischer-Witholt, S.J.; Guigard, E.; Kay, C.M.; Lemieux, M.J.

Oligomeric state study of prokaryotic rhomboid proteases

Biochim. Biophys. Acta

1818

3090-3097

2012

Escherichia coli, Bacillus spizizenii (E0U436), Haemophilus influenzae (P44783), Haemophilus influenzae, Escherichia coli DH5alpha, Bacillus spizizenii ATCC 23059 (E0U436), Haemophilus influenzae ATCC 51907 (P44783)

22921757

Arutyunova, E.; Panwar, P.; Skiba, P.M.; Gale, N.; Mak, M.W.; Lemieux, M.J.

Allosteric regulation of rhomboid intramembrane proteolysis

EMBO J.

33

1869-1881

2014

Escherichia coli, Haemophilus influenzae (P44783), Haemophilus influenzae, Providencia stuartii (P46116), Haemophilus influenzae ATCC 51907 (P44783)

25009246

Zoll, S.; Stanchev, S.; Began, J.; Skerle, J.; Lep?ik, M.; Peclinovska, L.; Majer, P.; Strisovsky, K.

Substrate binding and specificity of rhomboid intramembrane protease revealed by substrate-peptide complex structures

EMBO J.

33

2408-2421

2014

Escherichia coli

25216680

732230

Uritsky, N.; Shokhen, M.; Albeck, A.

The catalytic machinery of rhomboid proteases: Combined MD and QM simulations

J. Chem. Theory Comput.

8

4663-4671

2012

Escherichia coli (P09391), Escherichia coli

26605622

Lazareno-Saez, C.; Arutyunova, E.; Coquelle, N.; Lemieux, M.J.

Domain swapping in the cytoplasmic domain of the Escherichia coli rhomboid protease

J. Mol. Biol.

425

1127-1142

2013

Escherichia coli (P09391), Escherichia coli

23353827

732381

Reddy, T.; Rainey, J.K.

Multifaceted substrate capture scheme of a rhomboid protease

J. Phys. Chem. B

116

8942-8954

2012

Escherichia coli

22770371

Wolf, E.V.; Zeissler, A.; Vosyka, O.; Zeiler, E.; Sieber, S.; Verhelst, S.H.

A new class of rhomboid protease inhibitors discovered by activity-based fluorescence polarization

PLoS ONE

8

e72307

2013

Escherichia coli (P09391), Escherichia coli

23991088

732886

Foo, A.C.; Harvey, B.G.; Metz, J.J.; Goto, N.K.

Influence of hydrophobic mismatch on the catalytic activity of Escherichia coli GlpG rhomboid protease

Protein Sci.

24

464-473

2015

Escherichia coli

25307614

Zhou, Y.; Moin, S.M.; Urban, S.; Zhang, Y.

An internal water-retention site in the rhomboid intramembrane protease GlpG ensures catalytic efficiency

Structure

20

1255-1263

2012

Escherichia coli (P09391), Escherichia coli

22705210

Vinothkumar, K.R.; Pierrat, O.A.; Large, J.M.; Freeman, M.

Structure of rhomboid protease in complex with beta-lactam inhibitors defines the S2 cavity

Structure

21

1051-1058

2013

Escherichia coli (P09391), Escherichia coli

23665170

Wolf, E.V.; Zeissler, A.; Verhelst, S.H.

Inhibitor fingerprinting of rhomboid proteases by activity-based protein profiling reveals inhibitor selectivity and rhomboid autoprocessing

ACS Chem. Biol.

10

2325-2333

2015

Vibrio cholerae, Escherichia coli (P09391), Escherichia coli, Providencia stuartii (P46116)

26218717

753003

Yang, J.; Barniol-Xicota, M.; Nguyen, M.T.N.; Ticha, A.; Strisovsky, K.; Verhelst, S.H.L.

Benzoxazin-4-ones as novel, easily accessible inhibitors for rhomboid proteases

Bioorg. Med. Chem. Lett.

28

1423-1427

2018

Bacillus subtilis, Escherichia coli

29506958

753260

Wolf, E.V.; Seybold, M.; Hadravova, R.; Strisovsky, K.; Verhelst, S.H.

Activity-based protein profiling of rhomboid proteases in liposomes

ChemBioChem

16

1616-1621

2015

Escherichia coli

26032951

753975

Barniol-Xicota, M.; Verhelst, S.H.L.

Stable and functional rhomboid proteases in lipid nanodiscs by using diisobutylene/maleic acid copolymers

J. Am. Chem. Soc.

140

14557-14561

2018

Escherichia coli (P09391)

30347979

754206

Zhou, H.; Yu, H.; Zhao, X.; Yang, L.; Huang, X.

Molecular dynamics simulations investigate the pathway of substrate entry active site of rhomboid protease

J. Biomol. Struct. Dyn.

37

3445-3455

2018

Escherichia coli (P09391)

30175657