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Literature summary for 3.4.11.26 extracted from

  • Singh, R.; Goyal, V.D.; Kumar, A.; Sabharwal, N.S.; Makde, R.D.
    Crystal structures and biochemical analyses of intermediate cleavage peptidase role of dynamics in enzymatic function (2019), FEBS Lett., 593, 443-454 .
    View publication on PubMed
Search for more literature for 3.4.11.26

Cloned(Commentary)

Cloned (Comment) Organism
-
 Saccharomyces cerevisiae
expression in Escherichia coli BL21(DE3). Full-length Icp55 construct can not be expressed in the soluble fraction. Two truncated versions, Icp55t1 with 42 amino acid N-terminal truncation and Icp55t2 with 57 amino acid N-terminal truncation, are successfully expressed in soluble forms. All activity assays for Icp55 are reported for Icp55t1 construct Saccharomyces cerevisiae

Crystallization (Commentary)

Crystallization (Comment) Organism
free enzyme and in complex with inhibitor apstatin. The enzyme exists in a rapid equilibrium between monomer and dimer. The dimer, and not the monomer, is the active species with loop dynamics at the dimer interface playing an important role in activity Saccharomyces cerevisiae
oil microbatch method Saccharomyces cerevisiae

Protein Variants

Protein Variants Comment Organism
Y284D mutation of a conserved Tyr284 residue located at hinge helix that interacts with its counterpart across the dimer interface. Mutant is a monomer Saccharomyces cerevisiae

Inhibitors

Inhibitors Comment Organism Structure
apstatin crystal data in complex with apstatin Saccharomyces cerevisiae
EDTA complete loss of activity Saccharomyces cerevisiae
Tyr-Ala-Ala
-
 Saccharomyces cerevisiae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.72
-
 Tyr-Ala-Ala pH 7.5, 40°C Saccharomyces cerevisiae
1.8
-
 Tyr-Pro-Ala pH 7.5, 40°C Saccharomyces cerevisiae
2
-
 Met-Pro-Ala pH 7.5, 40°C Saccharomyces cerevisiae

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ reducing concentration of Mn2+ in reaction buffer from 1 mM to 6 microM reduces the activity of the enzyme by about 60% Saccharomyces cerevisiae
Mn2+ the activity of the enzyme depends critically on the presence of Mn2+. Reducing concentration of Mn2+ in reaction buffer from 1 mM to 0.006 mM reduces the activity of the enzyme by about 60%. Other divalent metal ions (Mg2+, Ca2+, Co2+, Ni2+ and Zn2+) fail to fully restore activity of the enzyme Saccharomyces cerevisiae
additional information not activating: Mg2+, Ca2+, Co2+, Ni2+, Zn2+ Saccharomyces cerevisiae

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
53000
-
 and 106000, gel filtration, truncated variant lacking 42 N-terminal amino acids Saccharomyces cerevisiae
106000
-
 dimer, gel filtration Saccharomyces cerevisiae
106000
-
 and 53000, gel filtration, truncated variant lacking 42 N-terminal amino acids Saccharomyces cerevisiae

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae P40051
-
-
Saccharomyces cerevisiae ATCC 204508 P40051
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Met-Pro-Ala + H2O
-
 Saccharomyces cerevisiae Met + Pro-Ala
-
 ?
additional information residue Tyr is preferred at P1 position. The enzyme additionally displays Xaa-Pro aminopeptidase activity in vitro Saccharomyces cerevisiae ?
-
 ?
additional information the enzyme is active towards substrates with proline at P1' position (M-/-PA and Y-/-PA). Icp55 cleaves off bulky residues from N-termini of proteins. Active towards substrates Y-/-AA, Y-/-TA and Y-/-SA Saccharomyces cerevisiae ?
-
 ?
additional information residue Tyr is preferred at P1 position. The enzyme additionally displays Xaa-Pro aminopeptidase activity in vitro Saccharomyces cerevisiae ATCC 204508 ?
-
 ?
Tyr-Ala-Ala + H2O
-
 Saccharomyces cerevisiae Tyr + Ala-Ala
-
 ?
Tyr-Pro-Ala + H2O
-
 Saccharomyces cerevisiae Tyr + Pro-Ala
-
 ?
Tyr-Ser-Ala + H2O
-
 Saccharomyces cerevisiae Tyr + Ser-Ala
-
 ?
Tyr-Thr-Ala + H2O
-
 Saccharomyces cerevisiae Tyr + Thr-Ala
-
 ?

Subunits

Subunits Comment Organism
dimer 2 * 52000, gel filtration, the enzyme exists in a rapid equilibrium between monomer and dimer. The dimer, not monomer, is the active species of the enzyme with loop dynamics at the dimer interface playing an important role in activity. Dynamics of Icp55 protein between two conformations of dimer are found to be important for activity of the enzyme Saccharomyces cerevisiae
monomer 1 * 52000, the enzyme exists in a rapid equilibrium between monomer and dimer. The dimer, not monomer, is the active species of the enzyme Saccharomyces cerevisiae
monomer and dimer, 1 * 58000 and 2 * 58000, calculated from sequence Saccharomyces cerevisiae

Synonyms

Synonyms Comment Organism
Icp55
-
 Saccharomyces cerevisiae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
 assay at Saccharomyces cerevisiae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.4
-
 Tyr-Ala-Ala pH 7.5, 40°C Saccharomyces cerevisiae
14
-
 Tyr-Pro-Ala pH 7.5, 40°C Saccharomyces cerevisiae
25.7
-
 Met-Pro-Ala pH 7.5, 40°C Saccharomyces cerevisiae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Saccharomyces cerevisiae

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
5
-
 Tyr-Ala-Ala pH 7.5, 40°C Saccharomyces cerevisiae

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.94
-
 Tyr-Ala-Ala pH 7.5, 40°C Saccharomyces cerevisiae
7.78
-
 Tyr-Pro-Ala pH 7.5, 40°C Saccharomyces cerevisiae
12.8
-
 Met-Pro-Ala pH 7.5, 40°C Saccharomyces cerevisiae