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Related Experiment Video

Updated: Jul 10, 2026

The Determination of Protease Specificity in Mouse Tissue Extracts by MALDI-TOF Mass Spectrometry: Manipulating PH to Cause Specificity Changes

The Determination of Protease Specificity in Mouse Tissue Extracts by MALDI-TOF Mass Spectrometry: Manipulating PH to Cause Specificity Changes

Published on: May 25, 2018

Structural basis for elastolytic substrate specificity in rodent alpha-chymases.

Jukka Kervinen¹, Marta Abad¹, Carl Crysler¹

¹Johnson & Johnson Pharmaceutical Research and Development, Structural Biology, Exton, Pennsylvania 19341.

The Journal of Biological Chemistry

|November 6, 2007

Summary

This summary is machine-generated.

Hamster chymase-2 (HAM2) exhibits unique elastolytic specificity due to its narrow S1 binding pocket, unlike hamster chymase-1 (HAM1). This divergence in enzyme activity is explained by specific amino acid changes, aiding prediction of chymase-like sequences.

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Defining Substrate Specificities for Lipase and Phospholipase Candidates

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Defining Substrate Specificities for Lipase and Phospholipase Candidates

Defining Substrate Specificities for Lipase and Phospholipase Candidates

Published on: November 23, 2016

Area of Science:

Enzymology
Structural Biology
Evolutionary Biology

Background:

Enzyme specificity evolves through changes in substrate binding sites.
Rodent alpha-chymases provide a model for studying enzyme evolution.
Understanding specificity divergence is key to predicting new enzyme functions.

Purpose of the Study:

To elucidate the structural basis for the atypical substrate specificity of hamster chymase-2 (HAM2).
To compare the enzymatic and structural properties of HAM2 with hamster chymase-1 (HAM1).
To identify key residues and sequence motifs predictive of elastolytic specificity in rodent alpha-chymases.

Main Methods:

Enzymatic characterization of HAM1 and HAM2.
X-ray crystallography of HAM2 complexed with a peptidyl inhibitor.
Analysis of substrate binding pocket (S1) dimensions and residue composition.
Phylogenetic comparison of chymase sequences.

Main Results:

HAM2 displayed elastolytic specificity (cleaving after Ala/Val), contrasting with HAM1's chymotryptic activity.
The crystal structure of HAM2 revealed a narrow, shallow S1 pocket accommodating small hydrophobic residues.
Four key residues in the S1 pocket (189, 190, 216, 226) explain the specificity difference between HAM1 and HAM2.
A conserved triplet (Asn189, Val190, Val216) in rodent alpha-chymases predicts elastolytic activity.
Guinea pig and rabbit chymases are the closest orthologs to rodent alpha-chymases.

Conclusions:

Specific amino acid substitutions in the S1 pocket dictate substrate specificity divergence in alpha-chymases.
The identified residue triplet serves as a reliable marker for predicting elastolytic specificity in novel chymase sequences.
This study provides a structural and evolutionary framework for understanding enzyme adaptation.