Subtilisin Enzymes

1. Front Matter

   Pages i-ix

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2. Structural Studies of Subtilisin Enzymes Alone and in Complex with Inhibitors

   1. The Complex Between Mesentericopeptidase and Eglin-C

      • Zbigniew Dauter, Christian Betzel, Nicolay Genov, Keith S. Wilson

      Pages 1-4

   2. Crystallographic Study of Eglin-C Binding to Thermitase

      • Alexei Teplyakov, Piet Gros, Wim G. J. Hol

      Pages 5-9

   3. Design of Specific Peptide Structures and Subtilisin Enzyme Inhibitors Using α, β-Dehydro-Residues

      • T. P. Singh, B. Padmanabhan, P. Narula, A. K. Saxena, C. Betzel, P. Sharma et al.

      Pages 11-20

   4. Structure and Function of Subtilisin BPN’ as Studied Through Crystallographic Studies on a Series of its Complexes with Genetically Engineered Proteinaceous Inhibitor SSI

      • T. Nonaka, T. Suzuki, N. Tanaka, S. Saito, T. Senda, K. Miura et al.

      Pages 21-27

   5. Three Dimensional Structure of the Antibiotic Bacitracin a Complexed to Two Different Subtilisin Proteases: Novel Mode of Enzyme Inhibition

      • Sabine Pfeffer-Hennig, Zbigniew Dauter, Michael Hennig, Wolfgang Höhne, Keith Wilson, Christian Betzel

      Pages 29-41

   6. Active Site Binding Loop Stabilization in the Subtilisin Inhibitor Eglin C: Structural and Functional Studies on Specifically Designed Mutants in Complex with Subtilisin and the Uncomplexed Inhibitor

      • Karsten Hipler, John P. Priestle, Joseph Rahuel, Markus G. Grütter

      Pages 43-47

   7. Crystal Structures of the Alkaline Proteases Savinase and Esperase from Bacillus Lentus

      • Christian Betzel, Silke Klupsch, Sven Branner, Keith S. Wilson

      Pages 49-61

3. New Enzymes

   1. Modelling and Engineering of Enzyme/Substrate Interactions in Subtilisin-Like Enzymes of Unknown 3-Dimensional Structure

      • Roland J. Siezen

      Pages 63-73

   2. Subtilases: Subtilisin-Like Serine Proteases

      • Roland J. Siezen

      Pages 75-93

   3. Cloning and Expression of the Gene Encoding a Novel Proteinase from TritiraChium Album Limber

      • Babru Sama, Tom Boone, Barbara Karan, Kenneth Chen, Raj Sachdev, Tsutomo Arakawa

      Pages 95-104

4. Structure-Function Studies

   1. Studies of Binding Sites in the Subtilisin from Bacillus Lentus by Means of Site Directed Mutagenesis and Kinetic Investigations

      • Hanne Grøn, Lene M. Bech, Steen Bech Sørensen, Morten Meldal, Klaus Breddam

      Pages 105-112

   2. Laundry Performance Of Subtilisin Proteases

      • A. M. Wolff, M. S. Showell, M. G. Venegas, B. L. Barnett, W. C. Wertz

      Pages 113-120

   3. Free Energy Perturbation Techniques Applied To Subtilisin BPN’ Stability

      • B. L. Barnett, Cathleen B. Turner

      Pages 121-131

5. Biophysical Probes and Mutagenesis

   1. Thermitase — Kinetic Differentiation to the Subtilisins

      • K. Peters, D. Brömine, G. Jahreis, S. Fittkau

      Pages 133-140

   2. Fluorescence Decay of Tryptophans in Serine Proteinases from Microorganisms: Relation to X-Ray Models

      • Nicolay Genov, Peter Nikolov, Christian Betzel, Keith Wilson

      Pages 141-145

   3. Propeptide-Mediated Folding in Subtilisin: The Intramolecular Chaperone Concept

      • Ujwal Shinde, Masayori Inouye

      Pages 147-154

   4. Roles of The N- and C- Terminal Pro-Sequences of Aqualysin I Precursor in the Processing and Extracellular Secretion of the Enzyme

      • Young-Choon Lee, Ichiro Terada, Takahisa Ohta, Hiroshi Matsuzawa

      Pages 155-157

   5. Crystal Structure Analysis of Subtilisin BPN’ Mutants Engineered for Studying Thermal Stability

      • Gary L. Gilliland, D. Travis Gallagher, Patrick Alexander, Philip Bryan

      Pages 159-169

   6. Random Mutagenesis of the Weak Calcium Binding Site in Subtilisin Carlsberg and Screening for Thermostability by Temperature-Gradient Gel Electrophoresis

      • Andrea Sättler, Susanne Kanka, Wolfgang Schrörs, Detlev Riesner

      Pages 171-182

Subtilisin is the most extensively studied model system for protein engineering. The primary motivating factor for the interest in subtilisin is the commercial utility of this class of proteases. The subtilisin symposium was the first international meeting to bring together a large number of groups that have focused on the subtilisins and the subtilases-the protein superfamily of subtilisin-like enzymes. The results presented at the symposium are in this way a unique compendium of a broad spectrum of work largely focused on harnessing the potential of site-directed mutagenesis to understand and deliberately alter the function of these enzymes toward a desired end. This sort of protein engineering has been extremely successful in subtilisin, with many such "engineered" enzymes now widely used in commer­ cial enterprises. In this regard the experience derived from subtilisin does represent practical protein engineering. It is becoming clear that subtilisin represents a larger class of enzymes, the subtilases, that include many of the human pro hormone-converting enzymes. As international collabo­ rative efforts to sequence entire genomes continue, we can reasonably expect that additional members of the subtilase class will be encountered. Whenever interest in a member of this class of enzyme arises, the work on subtilisin will serve as a guide to the analysis for what in bacillus, fungi, and industry is an everyday workhorse enzyme.

• enzyme
• enzymes
• fungi
• genes
• protein
• protein engineering
• system

• Genecor International, Inc., South San Francisco, USA

  Richard Bott

• European Molecular Biology Laboratory, Hamburg, Germany

  Christian Betzel

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