1GBI

ALPHA-LYTIC PROTEASE WITH MET 190 REPLACED BY ALA AND GLY 216 REPLACED BY LEU COMPLEX WITH METHOXYSUCCINYL-ALA-ALA-PRO-PHENYLALANINE BORONIC ACID

Summary for 1GBI

• Entry DOI: 10.2210/pdb1gbi/pdb
• Related PRD ID: PRD_000315
• Descriptor: ALPHA-LYTIC PROTEASE, METHOXYSUCCINYL-ALA-ALA-PRO-PHENYLALANINE BORONIC ACID INHIBITOR, SULFATE ION, ... (4 entities in total)
• Functional Keywords: active-site mutation, serine proteinase, hydrolase-hydrolase inhibitor complex, hydrolase/hydrolase inhibitor
• Biological source: Lysobacter enzymogenes
• Total number of polymer chains: 2
• Total formula weight: 20581.61

Authors

Mace, J.E.,Agard, D.A. (deposition date: 1995-09-06, release date: 1996-01-29, Last modification date: 2021-11-03)

Primary citation

Mace, J.E.,Agard, D.A.
Kinetic and structural characterization of mutations of glycine 216 in alpha-lytic protease: a new target for engineering substrate specificity.
J.Mol.Biol., 254:720-736, 1995

PubMed Abstract: Gly216 in the active site of the broadly specific MA190 mutant of alpha-lytic protease has been found to be remarkably tolerant of amino acid substitutions. Side-chains as large as Trp can be accommodated within the substrate-binding pocket without abolishing catalysis, and have major effects upon the substrate specificity of the enzyme. Kinetic characterization of eleven enzymatically active mutants against a panel of eight substrates clearly revealed the functional consequences of the substitutions at position 216. To understand better the structural basis for their altered specificity, the GA216 + MA190 and GL216 + MA190 mutants have been crystallized both with and without a representative series of peptide boronic acid transition-state analog inhibitors. An empirical description and non-parametric statistical analysis of structural variation among these enzyme: inhibitor complexes is presented. The roles of active site plasticity and dynamics in alpha-lytic protease function and substrate preference are also addressed. The results strongly suggest that substrate specificity determination in alpha-lytic protease is a distributed property of the active site and substrate molecule.

PubMed: 7500345
DOI: 10.1006/jmbi.1995.0650

Experimental method

X-RAY DIFFRACTION (2.3 Å)