A Practical Guide To Protein Engineering

This textbook introduces readers in an accessible and engaging way to the nuts and bolts of protein expression and engineering. Various case studies illustrate each step from the early sequence searches in online databases over plasmid design and molecular cloning techniques to protein purification and characterization. Furthermore, readers are provided with practical tips to successfully pursue a career as a protein engineer. With protein engineering being a fundamental technique in almost all molecular biology labs, the book targets advanced undergraduates and graduate students working in molecular biology, biotechnology and related scientific fields.

Protein engineering is the rational modification or redesign of proteins using genetic engineering. Thus, it is now possible to modify enzyme specifics, remodel antibodies, and redesign many multi-domain proteins for therapeutic purposes. While the procedures for the introduction of mutations have become routine, predicting and understanding the effects of these mutations can be complicated. This volume provides a comprehensive guide to the methods used at every stage of the engineering process, from the choice of mutation strategy, through protein stability studies, to critical evaluations of mammalian, yeast, and bacterial host expression systems. Protein Engineering: A Practical Approach is the first practical guide to this fascinating mixture of molecular biology, protein structure analysis, computation, and biochemistry. It combines a thorough theoretical foundation with detailed protocols and will be invaluable to all research workers in the area, from graduate students to senior investigators.

This textbook introduces readers in an accessible and engaging way to the nuts and bolts of protein expression and engineering. Various case studies illustrate each step from the early sequence searches in online databases over plasmid design and molecular cloning techniques to protein purification and characterization. Furthermore, readers are provided with practical tips to successfully pursue a career as a protein engineer. With protein engineering being a fundamental technique in almost all molecular biology labs, the book targets advanced undergraduates and graduate students working in molecular biology, biotechnology and related scientific fields.

PROTEIN ENGINEERING Principles and Practice Edited by JEFFREY L. CLELAND CHARLES S. CRAIK Proteins are involved in every aspect of life-structure, motion, catalysis, recognition and regulation. Protein Engineering: Principles and Practice provides a basic framework for understanding both proteins and protein engineering. This comprehensive book covers general, yet essential knowledge required for successful protein engineering, including everything from the fundamentals to modifying existing proteins and developing new proteins. The book begins by introducing the main concepts of protein engineering, including: understanding protein conformation, comprehending the relationship between protein composition and structure, and potential methods for predicting a protein's conformation. Other major subjects addressed are: * Using different host cell expression systems to produce specific proteins * Protein folding * Structure and function of proteins in relation to drug design * Construction of synthetic metal binding sites in proteins * Manufacture of tissue plasminogen activator * Generation of therapeutic antibodies This broad range of topics provides a solid foundation in protein engineering and supplies readers with knowledge essential to the design and production of proteins. Of primary interest to protein scientists-both students and researchers, in academia as well as industry-Protein Engineering is also extremely useful to chemical engineers, protein chemists, biochemists, and pharmaceutical chemists.

Experimental protein engineering and computational protein design are broad but complementary strategies for developing proteins with altered or novel structural properties and biological functions. By describing cutting-edge advances in both of these fields, Protein Engineering and Design aims to cultivate a synergistic approach to protein science

Why a Second Edition? The Second Edition provides practical answers to the general question, "How can I obtain useful sequence information from my protein or peptide?" rather than the more specific question asked in the first edition, "How can I obtain the N-terminal sequence?" Important new methods include ways of dealing with blocked N termini, computer analysis of protein sequences, and the recent revolution in mass spectrometry. Mass spectrophotometric characterization of proteins and peptides N-terminal sequencing of proteins with blocked N termini Internal amino acid sequence analysis after protease digestion in-gel and on-blot Improved microscale peptide purification methods Computer analysis of protein sequences New protocols tested and refined through everyday use in authors' laboratories Updated reference chapter covering all aspects of protein microsequencing

The desire to understand protein structure has been given new impetus by the explosive growth of biotechnology and the important role of proteins, natural and modified, in this technology. Protein molecules are machines, and protein engineering is opening up a whole new world of machinery on the molecular scale. This work is a simple, but in many ways detailed, introduction, to current knowledge, techniques, and applications. The volume is essentially in two parts, the first half covering a basic introduction to proteins appropriate at the undergraduate and early postgraduate level, which will prove a valuable teaching aid. The second half is a more advanced guide to concepts and methods, covering a range of aspects not previously collected in one volume. It will serve as a background reader and guide for advanced research study.

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