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Structural basis for differences in substrate selectivity in Kex2 and furin protein convertases
Todd Holyoak1, Charles A Kettner, Gregory A Petsko
1Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, Massachusetts 02454, USA.
Biochemistry
|March 3, 2004
Summary
The crystal structure of yeast Kex2 protease reveals insights into substrate specificity differences compared to its mammalian counterpart, furin. This study also proposes a mechanism for Kex2 activation by potassium ions.
Area of Science:
- Biochemistry
- Structural Biology
- Enzymology
Background:
- Kex2 is a yeast serine protease, a model for a family cleaving substrates at paired basic sites.
- Understanding Kex2 structure-function relationships is crucial for protease research.
Purpose of the Study:
- To determine the crystal structure of yeast Kex2 (ssKex2) in complex with a peptidyl boronic acid inhibitor.
- To elucidate structural differences in substrate recognition between Kex2 and mammalian furin.
- To identify cation binding sites and propose a mechanism for Kex2 activation.
Main Methods:
- X-ray crystallography at 2.2 A resolution.
- Complex formation between ssKex2 and Ac-Arg-Glu-Lys-Arg peptidyl boronic acid inhibitor.
- Comparative structural analysis with the furin homologue.
Main Results:
- The 2.2 A crystal structure of ssKex2-inhibitor complex was solved.
- Structural basis for P(2) and P(4) substrate recognition differences between Kex2 and furin was proposed.
- Structural rationale for Kex2's lack of P(6) recognition was provided.
- Monovalent cation binding sites were identified, including potassium.
Conclusions:
- The study provides a structural understanding of Kex2 substrate specificity.
- A mechanism for Kex2 activation by potassium ions is proposed.
- Structural insights advance the study of Kex2 and related proteases.