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Electrostatics of mesophilic and psychrophilic trypsin isoenzymes: Qualitative evaluation of electrostatic differences at the substrate binding site
Author(s) -
Gorfe Alemayehu A.,
Brandsdal Bjørn O.,
Leiros HannaKirsti S.,
Helland Ronny,
Smalås Arne O.
Publication year - 2000
Publication title -
proteins: structure, function, and bioinformatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.699
H-Index - 191
eISSN - 1097-0134
pISSN - 0887-3585
DOI - 10.1002/(sici)1097-0134(20000801)40:2<207::aid-prot40>3.0.co;2-u
Subject(s) - psychrophile , electrostatics , trypsin , substrate (aquarium) , chemistry , mesophile , isozyme , enzyme , biochemistry , biology , ecology , bacteria , genetics
A qualitative evaluation of electrostatic features of the substrate binding region of seven isoenzymes of trypsin has been performed by using the continuum electrostatic model for the solution of the Poisson‐Boltzmann equation. The sources of the electrostatic differences among the trypsins have been sought by comparative calculations on selective charges: all charges, conserved charges, partial charges, unique cold trypsin charges, and a number of charge mutations. As expected, most of the negative potential at the S 1 region of all trypsins is generated from Asp 189 , but the potential varies significantly among the seven trypsin isoenzymes. The three cold active enzymes included in this study possess a notably lower potential at and around the S 1 ‐pocket compared with the warm active counterparts; this finding may be the main contribution to the increased binding affinity. The source of the differences are nonconserved charged residues outside the specificity pocket, producing electric fields at the S 1 ‐pocket that are different in both sign and magnitude. The surface charges of the mesophilic trypsins generally induce the S 1 pocket positively, whereas surface charges of the cold trypsins produce a negative electric field of this region. Calculations on mutants, where charged amino acids were substituted between the trypsins, showed that mutations in Loop2 (residues 221B and 224) and residue 175, in particular, were responsible for the low potential of the cold enzymes. Proteins 2000;40:207–217. © 2000 Wiley‐Liss, Inc.