Any feedback?
Please rate this page
(enzyme.php)
(0/150)

BRENDA support

BRENDA Home
show all | hide all No of entries

Information on EC 3.4.21.88 - Repressor LexA and Organism(s) Mycobacterium tuberculosis and UniProt Accession P9WHR7

for references in articles please use BRENDA:EC3.4.21.88
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
     3 Hydrolases
         3.4 Acting on peptide bonds (peptidases)
             3.4.21 Serine endopeptidases
                3.4.21.88 Repressor LexA
Specify your search results
Select one or more organisms in this record: ?
This record set is specific for:
Mycobacterium tuberculosis
UNIPROT: P9WHR7 not found.
Show additional data
Do not include text mining results
Include (text mining) results
Include results (AMENDA + additional results, but less precise)
Word Map
hide
The taxonomic range for the selected organisms is: Mycobacterium tuberculosis
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
hide
Hydrolysis of Ala84-/-Gly bond in repressor LexA
Synonyms
lexa repressor, lexa protein, lexa1, repressor lexa, lexa transcriptional repressor, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LexA repressor
-
-
-
-
SOS regulatory protein dinR
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
84721-00-6
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
NaCl
variation of the sodium chloride concentration from 25 mM to 1.5 M do not show any significant change in the autoproteolysis of MtLexA
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
9.5 - 11
self-cleavage activity at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37 - 55
self-cleavage activity at
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
LexA is a critical protein involved in the bacterial SOS response, which consists of the coordinated activation of a network of genes required for DNA repair and mutagenesis in response to DNA damage. The clinical relevance of the SOS response in bacteria can be attributed not only to its involvement in virulence and mutagenesis, but also in the spread of antibiotic resistance
additional information
a complex of Mycobacterium tuberculosis LexA and the cognate SOS box is modeled in which the mutual orientation of the two N-terminal domains differs from that in the Escherichia coli LexA-DNA complex, complex structure analysis, overview
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
27600
Western blotting, Myc-tagged LexA
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
2 * 12000, C-terminal catalytic core, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
the autocatalytic cleavage of MtLexA and the mutant proteins is analyzed, effects of pH and temperature, the enzyme is autocatalytically active at pH 9.5-11.0 and at up to 55°C
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystallization of C-terminal domain, space groups P3221 and P31, to 2.9 and 2.75 A resolution, respectively
purified recombinant His-tagged wild-type LexA and mutant enzymes S160A, K197A, and G126D, the isolated C-terminal segment and the N-domain, wild-type LexA diffraction-quality crystals grow by vapour diffusion method in about three months from a drop consisting of 0.002 ml of 6 mg/ml protein solution and 0.002 ml of 100 mM bis-Tris, pH 6.5 or pH 8.5, 100 mM NaCl or 200 mM MgCl2, 5% glycerol, 25% PEG 3350 (form I and form II, respectively). The microbatch-under-oil method is used for crystallization of the C-domain by mixing of 0.002 ml of 10 mg/ml protein solution with 0.002 ml 100 mM Tris-HCl, pH 7.5, 25% glycerol, and 40% v/v pentaerythritol ethoxylate (15/4 EO/OH). Diffraction-quality crystals of the S160A mutant are obtained by microbatch-under-oil method and mixing of 0.002 ml 10 mg/ml protein and 0.002 ml 100 mM bis-Tris, pH 6.5, 200 mM MgCl2, and 25% w/v PEG 3350 (form III), and of mutant K197A by mixing of 0.002 ml 10 mg/ml protein solution with 0.002 ml crystallization solution containing 100 mM HEPES, pH 7.5, 20 mM MgCl2, and 22% w/v polyacrylic acid sodium salt (form IV), while crystals of the G126D mutant are grown from a solution consisting of 0.002 ml of 10 mg/ml protein with 0.002 ml of 100 mM Tris-HCl, pH 8.5, 100 mM sodium acetate trihydrate, and 30% w/v PEG 4000 (form V), mutant crystals grow about 2 months, X-ray diffraction structure determination and analysis at 1.48-2.25 A resolution
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
G126D
site-directed mutagenesis, cleavage site mutant
K197A
site-directed mutagenesis, active site mutant
S160A
site-directed mutagenesis, active site mutant
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
55
above, melting temperature of wild-type and mutant enzymes
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged wild-type and mutant enzymes, as well as enzyme fragments from Escherichia coli strain BL21(DE3) pLysS by nickel affinity chromatography and gel filtration, to over 95% purity
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
gene lexA, recombinant expression of His-tagged wild-type LexA and mutant enzymes S160A, K197A, and G126D, the isolated C-terminal segment and the N-domain in Escherichia coli strain BL21(DE3) pLysS
the lexA gene, including its promoter region, cloned into pEJMyc to give pKS04, giving LexA with an in-frame C-terminal Myc tag. Plasmids pKS04, pKS04mut1 (one residue deleted between the possible start codons) and the pEJMyc vector transformed separately into Mycobacterium smegmatis strain mc2155
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Smollett, K.L.; Fivian-Hughes, A.S.; Smith, J.E.; Chang, A.; Rao, T.; Davis, E.O.
Experimental determination of translational start sites resolves uncertainties in genomic open reading frame predictions - application to Mycobacterium tuberculosis
Microbiology
155
186-197
2009
Mycobacterium tuberculosis (P9WHR7), Mycobacterium tuberculosis H37Rv (P9WHR7)
Manually annotated by BRENDA team
Chandran, A.; Prabu, J.; Manjunath, G.; Patil, K.; Muniyappa, K.; Vijayan, M.
Crystallization and preliminary X-ray studies of the C-terminal domain of Mycobacterium tuberculosis LexA
Acta Crystallogr. Sect. F
66
1093-1095
2010
Mycobacterium tuberculosis (P9WHR7), Mycobacterium tuberculosis H37Rv (P9WHR7)
Manually annotated by BRENDA team
Smollett, K.L.; Smith, K.M.; Kahramanoglou, C.; Arnvig, K.B.; Buxton, R.S.; Davis, E.O.
Global analysis of the regulon of the transcriptional repressor LexA, a key component of SOS response in Mycobacterium tuberculosis
J. Biol. Chem.
287
22004-22014
2012
Mycobacterium tuberculosis (P9WHR7), Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv (P9WHR7)
Manually annotated by BRENDA team
Chandran, A.; Srikalaivani, R.; Paul, A.; Vijayan, M.
Biochemical characterization of Mycobacterium tuberculosis LexA and structural studies of its C-terminal segment
Acta Crystallogr. Sect. D
75
41-55
2019
Mycobacterium tuberculosis (P9WHR7), Mycobacterium tuberculosis H37Rv (P9WHR7), Mycobacterium tuberculosis ATCC 25618 (P9WHR7)
Manually annotated by BRENDA team