1NLU

Pseudomonas sedolisin (serine-carboxyl proteinase) complexed with two molecules of pseudo-iodotyrostatin

Summary for 1NLU

• Entry DOI: 10.2210/pdb1nlu/pdb
• Related: 1ga6 1kdv
• Related PRD ID: PRD_000754
• Descriptor: SEDOLISIN, PSEUDO-IODOTYROSTATIN, CALCIUM ION, ... (4 entities in total)
• Functional Keywords: pscp, hydrolase-hydrolase inhibitor complex, hydrolase/hydrolase inhibitor
• Biological source: Pseudomonas sp.; More
• Cellular location: Periplasm: P42790
• Total number of polymer chains: 3
• Total formula weight: 39335.83

Authors

Wlodawer, A.,Li, M.,Gustchina, A.,Dauter, Z.,Uchida, K.,Oyama, H.,Glodfarb, N.E.,Dunn, B.M.,Oda, K. (deposition date: 2003-01-07, release date: 2004-01-20, Last modification date: 2012-12-12)

Primary citation

Wlodawer, A.,Li, M.,Gustchina, A.,Oyama, H.,Oda, K.,Beyer, B.B.,Clemente, J.,Dunn, B.M.
Two inhibitor molecules bound in the active site of Pseudomonas sedolisin: a model for the bi-product complex following cleavage of a peptide substrate.
Biochem.Biophys.Res.Commun., 314:638-645, 2004

PubMed Abstract: High-resolution crystallographic analysis of a complex of the serine-carboxyl proteinase sedolisin with pseudo-iodotyrostatin revealed two molecules of this inhibitor bound in the active site of the enzyme, marking subsites from S3 to S3('). The mode of binding represents two products of the proteolytic reaction. Substrate specificity of sedolisin was investigated using peptide libraries and a new peptide substrate for sedolisin, MCA-Lys-Pro-Pro-Leu-Glu#Tyr-Arg-Leu-Gly-Lys(DNP)-Gly, was synthesized based on the results of the enzymatic and crystallographic studies and was shown to be efficiently cleaved by the enzyme. The kinetic parameters for the substrate, measured by the increase in fluorescence upon relief of quenching, were: k(cat)=73+/-5 s(-1), K(m)=0.12+/-0.011 microM, and k(cat)/K(m)=608+/-85 s(-1)microM(-1).

PubMed: 14733955
DOI: 10.1016/j.bbrc.2003.12.130

Experimental method

X-RAY DIFFRACTION (1.3 Å)