Laboratory Techniques In Membrane Biophysics

The present manual contains a collection of laboratory instructions used during an international training course on membrane biophysics which was held at Homburg in the fall of 1966. The selection of the topics dealt with in the various chapters depended on the scientific interest of the available teachers and on the availability of the necessary equipment in our laboratories. Thus, the material included in this volume does not add up to a systematic course in membrane biophysics. Instead it represents a more fortuitous collection of laboratory problems. In addition, some authors place more emphasis on teaching the more technical aspects of a method whereas others are primarily concerned with the demonstra tion of a significant biological phenomenon. Nevertheless, in spite of such differences of emphasis and a somewhat haphazard choice of a few methods and phenomena among many others of similar importance, it was felt that the publication of the material is desirable. Since no other laboratory manual exists so far, the present laboratory problems which were tested in actual practice may serve as a useful basis for the shaping of further training courses or for laboratory courses for graduate students in biophysics and physiology. Our thanks are due to the authors and the publisher who were patient and kind enough to cooperate with the editors during the long period between the end of the course and the appearance of the book.

The present manual contains a collection of laboratory instructions used during an international training course on membrane biophysics which was held at Homburg in the fall of 1966. The selection of the topics dealt with in the various chapters depended on the scientific interest of the available teachers and on the availability of the necessary equipment in our laboratories. Thus, the material included in this volume does not add up to a systematic course in membrane biophysics. Instead it represents a more fortuitous collection of laboratory problems. In addition, some authors place more emphasis on teaching the more technical aspects of a method whereas others are primarily concerned with the demonstra tion of a significant biological phenomenon. Nevertheless, in spite of such differences of emphasis and a somewhat haphazard choice of a few methods and phenomena among many others of similar importance, it was felt that the publication of the material is desirable. Since no other laboratory manual exists so far, the present laboratory problems which were tested in actual practice may serve as a useful basis for the shaping of further training courses or for laboratory courses for graduate students in biophysics and physiology. Our thanks are due to the authors and the publisher who were patient and kind enough to cooperate with the editors during the long period between the end of the course and the appearance of the book.

This volume provides recent advances in the field of biophysics of membrane proteins. Chapters are divided into several parts: detailing biochemistry and functional analysis, experimental and theoretical structural determinations, membrane protein dynamics, and conformation studies. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Biophysics of Membrane Proteins: Methods and Protocols aims to provide comprehensive protocols with notes to help further the understanding of key membrane protein structure and function for students, academics, and industrial researchers.

This manual collects in the form of laboratory protocols a series of experiments in the field of Membrane Transport and Membrane Bioenergetics. It represents the experience accumulated during four advanced courses held at the Depart ment of Biochemistry of the Swiss Federal Institute of Technology on behalf of Federation of European Biochemical Societies (FEBS) in the years 1975 through 1978. The idea of collecting the experiments into a laboratory manual developed as a response to a demand from the students who took part in the courses. Further motivation came with the fmding that, in planning the laboratory sessions, the teaching staff had no organized, modern source of information in the literature. The experiments presented cover most areas of importance in the subject mat ter. Their presentation has been continuously modified in the course of the four years during which the manual took shape, to accommodate to experience and various suggestions. In their present form, all of the experiments described have been repeatedly practiced to optimize their execution. Efforts have been made to combine in the manual classical experiments, and techniques which require relatively unsophisticated instrumentation and can therefore be carried out in most laboratories, with more modern experiments and relatively newer technol ogies. In its present form, the manual should therefore provide a usefui tool in the hands of researchers and laboratory teachers at different levels of sophisti cation and instrumentation.

This volume provides recent advances in the field of biophysics of membrane proteins. Chapters are divided into several parts: detailing biochemistry and functional analysis, experimental and theoretical structural determinations, membrane protein dynamics, and conformation studies. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Biophysics of Membrane Proteins: Methods and Protocols aims to provide comprehensive protocols with notes to help further the understanding of key membrane protein structure and function for students, academics, and industrial researchers.

Physics, mathematics and chemistry all play a vital role in understanding the true nature and functioning of biological membranes, key elements of living processes. Besides simple spectroscopic observations and electrical measurements of membranes we address in this book the phenomena of coexistence and independent existence of different membrane components using various theoretical approaches. This treatment will be helpful for readers who want to understand biological processes by applying both simple observations and fundamental scientific analysis. It provides a deep understanding of the causes and effects of processes inside membranes, and will thus eventually open new doors for high-level pharmaceutical approaches towards fighting membrane- and cell-related diseases.

The purposes of this senes were discussed in the preface to Volume I: to present "a range of methods . . . from the physical to the physiological . . . in sufficient detail for the reader to use them in his laboratory" and also to describe "the theoretical backgrounds of the methods and their limita tions in membrane biology" so that the reader will be enabled "to evaluate more critically and to understand more fully data obtained by methods foreign to [his] usual experiences. " The chapter by Lee, Birdsall, and Metcalfe with which Volume 2 begins accomplishes these twin goals with a thorough description of the application of nuclear magnetic relaxation measurements to membrane biology together with a lucid and succinct integration of the results of such studies into present concepts of the organi zation of membrane lipids. This then permits speculation on the physical basis of membrane permeability. The powerful tool of NMR spectroscopy will have even fuller application with the development of techniques, al ready partially exploited, for l3C-Iabeling of specific carbon atoms in lipid molecules and with extension of the observations to membrane proteins. The following two chapters, by Glick and by Laine, Stellner, and Hako mori, describe the isolation and characterization of membrane glycoproteins and membrane glycolipids, respectively.