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Subcellular particles, structures and organelles
Immum,Jwmi,tr~. 1'4"6, ~, ,~i [3. pp. 91 ~44
Pcrg,trntm Pr¢-,
Pr{ntcd in G r e a t B r l t , d n
BOOK REVIEXVS Subcellular Particles. Structures and Organdies---Edited by' Allen I. Laskin and Jerold A. Last. Marcel Dekker. New York. 1974. 305pp. 524.75. This volume is one in a continuing series of Methods in 3,1olecuk," Bioloqy. and it is in fact Volume 5 in the series. The particular emphasis of this volume and of this series is on methodology, and how to work v, ith subcellular particles and organelles. This is a particularly useful lield to review, summarize, and extract the good from the bad because there are so many ways to try to isolate so many subcellular organelles. The first chapter in this volume by Gary R. Craven. Laboratory of Molecular Biology. University of Wisconsin. concerns "'The Structure of the Bacterial Ribosome". In line with the sense of the series. Craven starts out with the preparation of ribosomes, followed by the extraction and identification of the individual proteins of bacterial ribosomes. He then goes on to review our knowledge of how to reconstitute ribosomes from the purified individual components. The description of the processes of the preparation of ribosomal components and their reconstitution are adequate but not overwhelming. Nor, is there totally reconciliation between the procedures proposed by Craven and those of Nomura. An.',~.vay. it is a good review, and helpful to anyone that wishes to start into this field. The second chapter concerns "'The Purification of Bacterial Flagellae and their Components" by DePamphilis, also of The Department of Biochemistry of the University of Wisconsin. Bacterial flagetlae are a special breed of flagellae, and are generally held to be different in detail, if not in principal, from the flagellae and cilia of higher organisms. It turns out that the best way to prepare flagellae from bacteria, as B. subtilis, is to make spheroplasts and then to lyse these and purlS' the flagellae by ammonium sulfate precipitation, differential centrifugation. and CsCI gradient centrifugation. The flagellae band in CsC1 at a density of 1,30 g,,ml and with a final yield of 40",{ of the original flagellae. Whether or not flagellae of bacteria grow from basal bodies as they do in eukaryote organisms, is a question discussed but not decided in the present review article. In any case, the protocols for the purification of flagellae are excellent. The review than goes on to discuss the separation of flagellae into component parts, flagellin, apparently a flagellar structural protein, which when separated into sub-units by extraction with either acidic or basic solutions reaggregate into filaments when returned to pH 7. The disassociation of filaments to flagellin results in residue flagellar hooks, which remain attached to pseudo-basal body complexes. The hook-basal body complex bands in Renografin at about 1.20 g'ml and is thus not very dense. The whole field of the study of the development, physical chemistry, and even the biochemistry of bacterial flagellae is clearly at its beginning, and represents a challenging field. The third chapter in this volume is by' Neville and Kahn, "'Isolation of Plasma Membranes for Cell Surface Membrane Receptor Studies". The idea is admirable. Let us isolate membranes from different kinds of mammalian cells; find out what is different between those of abnormal cells and normal ceils, and what is different between membrane receptors of cells, normal or transformed by oncogenic viral agents. Neville and Kahn educe rigorous criteria for the evaluation of membrane preparation, purity and homogeneity in several different ways, all good. They then outline all presently fashionable methods of membrane isolation including a very good literature review of
the subject. This is an excellent article for an investigator to use in starting any membrane receptor isolation endeavor. The appendix includes a biow-b}-blow account of how to get liver cell plasma membranes from mice. Chapter 4 concerns the "'Isolation of L.vsosomes'" by Gerald L. Rowin. Lysosomes have been known for twenty years since their discovery by deDuve. Lysosomes are very fragile. They are always prepared by differential centrifugation (in general they band above mitochondria), and they are invariably ruptured to some degree during their isolation. and therefore, release their bad enzymes which cause degradation of other tissue components. Rowin outlines all of the classical methods for isolation of lysosomes, and then goes on to treat in detail the observation that Triton WR-1339 cat, ses liver lysosomes of rats previously injected intravenously with this detergent, to become larger, less dense, and more easily separable from other subcellular components than is the case in the absence of the detergent. By the use of Triton WR-1339. lysosomes may be isolated in a form minimally contaminated by mitochondria. peroxisomes or microsomes. This is an excellent review for those who wish to find out how to isolate and study lysosomes. "Isolation of the Golgi Apparatus" is by William P. Cunningham. The Golgi apparatus is very difficult to prepare because it is so easily disrupted, and unluckily it is necessary to isolate the Golgi apparatus because it is so important. It plays a role in the concentration of secretory granules, in the attachment of carbohydrate moieties to proteins to form mucopolysaccharide and gl}coprotein secretory products, and it acts as a concentrating mechanism. Cunningham discusses the isolation of Golgi apparatus from epididymis, onion stem. rat liver, and rat testis. They are all difficult, but the techniques are all given, and it is possible to isolate the Golgi apparatus from all of these tissues. Anybody who really wants to isolate a Golgi apparatus himself, should read this article. It is complete. good. and really useful. It includes a complete review of the literature on Golgi iso[ation. The sixth chapter in this volume is on "'Zonal Centrifugation'" by Carl A. Price (my former undergraduate advisee) and it gives us good information about how to use zonal centrifuges. It is written in easy to understand terms. It gives us examples of different kinds of materials used for gradients, and of the kinds of separations to be expected. For newcomers to the field of zonal centrifugation, read this chapter, which includes a complete summary of all kinds of zonal rotors now available. Chapter 7 concerns "Methods in Electron Microscopy" by Breese and Zacharia. It includes examination of particles of ultrathin sections, of scanning electron microscopy, and related techniques. To quote the authors, "'studying electron microscope techniques in any text is not a substitute for actually performing the procedures in a laboratory with one's own hands". The reviewer believes this explicitly. Electron microscopy is best learned by apprenticeship. There are few principals that can be learned by reading a book, but these principles, such as they are, are all to be found in the article by Breese and Zacharia. The final chapter, "'Negative Staining for Electron Microscopy" is by DePamphilis. This again on Formvar coated grids, carbon film, and negative staining. Negative staining may have its place, but in this reviewer's experience. it is the cause of an unusual number of artifacts. 91
92
Book Re~iews
For those cases in which artit:acts are believed to not be induced by negative staining. DePamphilis's chapter is to be recommended. Overall. this is an excellent volume: full of information. lots of literature cited, lots of actually how to do-it-your-
self. and all in a nice small ~olume. which o n b costs 24.75 dollars. Dicision of Biology JAMES BONNER California Institute of Technology Pasadena. CA 91 i09. U.S.A.
Membrane Mediated Information--1. Biochemical Functions--Edited b.v P. W. Kent. American Elsevier, New York. 1973. 231 pp. 515.50. "'What's in a name?" or. indeed in a title'? The ~olume revie~cd hcrc contains a series of papers linked b? the common theme of study of oligosaccharide chains present at the surface of biological lipid membranes. These polymers are considered both in their own right, and as components of more complex and larger polymers, glycoproteins and glycolipids, Animal cells, especially cultured mammalian cell lines receive the greatest attention from the authors, though a three chapter section on viruses offers a view of membrane components of some enveloped viruses. "Information" is discussed in the opening article, by the late Richard Winzler. who points the number of permutations possible in a structure as simple as a disaccharide (16. in contrast to only 2 for a dipeptide). Many of the other papers tend to enlarge or detail for us the repertoire of oligosaccharide structure, but on the whole the readers are left to their own resources and to make their own inferences as to the "'informational" or biological meaning of structural variations in short sugar chains. (Important exceptions to them are the chapter on heterophile agglutinins and ABH blood group antigens dealing with systems in which both chemistry and immunology is understood). This reviewer appreciated such freedom to speculate, or perhaps ponder, and believes that the volume is valuable
as a special sort of extended review, of great use to biologists of all sorts, who study' the cell surface in terms of function, or serological specificity (but often in ignorance of chemical details) and conversely to chemists in search of functions and roles for particle structures. Most of the chapters combine brief reviews of an area with descriptions of fairly recent experimental work. Such combinations allow rapid entry into an area, e.g. glycolipid structure in transformed and non-transformed cells, together with a good appreciation of the technical aspects of work, the details of experiments too often omitted from more strictly review essays. Thus. though many of the experiments described here have been published in more extended lbrm elsewhere, they serve to reinforce the review aspects of the volume. Membrane glycolipids and glycoproteins, and some aspects of membrane phospholipids are dealt with in compact and adequate fashion. I would recommend the volume as what might be termed a "'working introduction" to an aspect of cell surface chemistry which, I predict will intrude itself more and more into cell biology.
Biology Department The Johns Hopkins Unicersirv Baltimore, MD 21218. U.S.A.
MICHAEL EDIDIN
Cellular Selection and Regulation in the Immune Response---Edited by Gerald M. Edelman. North Holland PuNishing Company, Amsterdam, 1974, 330 pp., S23.10. The collection of papers gathered together in this volume provide stimulating and interesting reading. A wide range of topics is covered in considerable detail there being 17 papers on the following themes: quantitation of clonal selection, immunogenetics, the cell surface and various aspects of lymphocyte stimulation, immunoglobulin synthesis and regulation of the immune response. It would be unfair to pick out articles as being good since the majority of articles are of a uniformly high standard and are a good combination of fact and speculation. One of the papers was. however, conspicuous in the absence of sufficient statistical qualification of the data presented in its numerous tables. Most of the information being presented in each article is not now new. but several of the papers each constitute a short review of their specified areas. They would therefore, provide a ready reference source for someone who quickly wants to be brought up to date with several different aspects of immunology. A few examples should give some idea of the scope covered in each section. Jerne in his paper 'Clonal Selection in a Lymphocyte Network' outlines the difficulties of quantitating clonal selection and discusses the possibilities of building a biologically acceptable mathematical model of the response of the immune response to antigens. Edelman in the same section considers the origin and mechanism of specificity in clonal selection b.~ covering
possible evolutionary relationships between histocompatibility loci. flz microglobulin and immunoglobulin; antibody specificity: the requirements of clonal selection and the ontogeny of specific antibody binding cells. The immunogenetics section includes articles on both histocompatibility linked genetic control of the immune response (McDevitt et al.) and the genetic fine structure of the H-2 complex (Shreffler). The former paper reviews and discusses evidence for and against the hypothesis that H-linked Ir genes are expressed only on T derived antigen reactive ceils. The latter paper contains a useful section for those unfamiliar with this field on the construction of the genetic map of the H-2 region of the, chromosome and describes the production, characteristics and potential uses of antisera to antigens controlled by genes localised in several papers but particular b that of Milstein et ,1., Histocompatibility antigens and other surface markers are used as tools to probe membrane structure. The binding of Ag-Ab complexes to cells and the possible role of this as a controlling function in the immune response is discussed. The proliferation and maturation of lymphocytes, the turnover of surface Ig, its site of synthesis and its relationship to secreted Ig are also covered in some depth. The various characteristics of Con A, its capacity to be both a stimulator and an inhibitor, its detailed structure
Pcrg,trntm Pr¢-,
Pr{ntcd in G r e a t B r l t , d n
BOOK REVIEXVS Subcellular Particles. Structures and Organdies---Edited by' Allen I. Laskin and Jerold A. Last. Marcel Dekker. New York. 1974. 305pp. 524.75. This volume is one in a continuing series of Methods in 3,1olecuk," Bioloqy. and it is in fact Volume 5 in the series. The particular emphasis of this volume and of this series is on methodology, and how to work v, ith subcellular particles and organelles. This is a particularly useful lield to review, summarize, and extract the good from the bad because there are so many ways to try to isolate so many subcellular organelles. The first chapter in this volume by Gary R. Craven. Laboratory of Molecular Biology. University of Wisconsin. concerns "'The Structure of the Bacterial Ribosome". In line with the sense of the series. Craven starts out with the preparation of ribosomes, followed by the extraction and identification of the individual proteins of bacterial ribosomes. He then goes on to review our knowledge of how to reconstitute ribosomes from the purified individual components. The description of the processes of the preparation of ribosomal components and their reconstitution are adequate but not overwhelming. Nor, is there totally reconciliation between the procedures proposed by Craven and those of Nomura. An.',~.vay. it is a good review, and helpful to anyone that wishes to start into this field. The second chapter concerns "'The Purification of Bacterial Flagellae and their Components" by DePamphilis, also of The Department of Biochemistry of the University of Wisconsin. Bacterial flagetlae are a special breed of flagellae, and are generally held to be different in detail, if not in principal, from the flagellae and cilia of higher organisms. It turns out that the best way to prepare flagellae from bacteria, as B. subtilis, is to make spheroplasts and then to lyse these and purlS' the flagellae by ammonium sulfate precipitation, differential centrifugation. and CsCI gradient centrifugation. The flagellae band in CsC1 at a density of 1,30 g,,ml and with a final yield of 40",{ of the original flagellae. Whether or not flagellae of bacteria grow from basal bodies as they do in eukaryote organisms, is a question discussed but not decided in the present review article. In any case, the protocols for the purification of flagellae are excellent. The review than goes on to discuss the separation of flagellae into component parts, flagellin, apparently a flagellar structural protein, which when separated into sub-units by extraction with either acidic or basic solutions reaggregate into filaments when returned to pH 7. The disassociation of filaments to flagellin results in residue flagellar hooks, which remain attached to pseudo-basal body complexes. The hook-basal body complex bands in Renografin at about 1.20 g'ml and is thus not very dense. The whole field of the study of the development, physical chemistry, and even the biochemistry of bacterial flagellae is clearly at its beginning, and represents a challenging field. The third chapter in this volume is by' Neville and Kahn, "'Isolation of Plasma Membranes for Cell Surface Membrane Receptor Studies". The idea is admirable. Let us isolate membranes from different kinds of mammalian cells; find out what is different between those of abnormal cells and normal ceils, and what is different between membrane receptors of cells, normal or transformed by oncogenic viral agents. Neville and Kahn educe rigorous criteria for the evaluation of membrane preparation, purity and homogeneity in several different ways, all good. They then outline all presently fashionable methods of membrane isolation including a very good literature review of
the subject. This is an excellent article for an investigator to use in starting any membrane receptor isolation endeavor. The appendix includes a biow-b}-blow account of how to get liver cell plasma membranes from mice. Chapter 4 concerns the "'Isolation of L.vsosomes'" by Gerald L. Rowin. Lysosomes have been known for twenty years since their discovery by deDuve. Lysosomes are very fragile. They are always prepared by differential centrifugation (in general they band above mitochondria), and they are invariably ruptured to some degree during their isolation. and therefore, release their bad enzymes which cause degradation of other tissue components. Rowin outlines all of the classical methods for isolation of lysosomes, and then goes on to treat in detail the observation that Triton WR-1339 cat, ses liver lysosomes of rats previously injected intravenously with this detergent, to become larger, less dense, and more easily separable from other subcellular components than is the case in the absence of the detergent. By the use of Triton WR-1339. lysosomes may be isolated in a form minimally contaminated by mitochondria. peroxisomes or microsomes. This is an excellent review for those who wish to find out how to isolate and study lysosomes. "Isolation of the Golgi Apparatus" is by William P. Cunningham. The Golgi apparatus is very difficult to prepare because it is so easily disrupted, and unluckily it is necessary to isolate the Golgi apparatus because it is so important. It plays a role in the concentration of secretory granules, in the attachment of carbohydrate moieties to proteins to form mucopolysaccharide and gl}coprotein secretory products, and it acts as a concentrating mechanism. Cunningham discusses the isolation of Golgi apparatus from epididymis, onion stem. rat liver, and rat testis. They are all difficult, but the techniques are all given, and it is possible to isolate the Golgi apparatus from all of these tissues. Anybody who really wants to isolate a Golgi apparatus himself, should read this article. It is complete. good. and really useful. It includes a complete review of the literature on Golgi iso[ation. The sixth chapter in this volume is on "'Zonal Centrifugation'" by Carl A. Price (my former undergraduate advisee) and it gives us good information about how to use zonal centrifuges. It is written in easy to understand terms. It gives us examples of different kinds of materials used for gradients, and of the kinds of separations to be expected. For newcomers to the field of zonal centrifugation, read this chapter, which includes a complete summary of all kinds of zonal rotors now available. Chapter 7 concerns "Methods in Electron Microscopy" by Breese and Zacharia. It includes examination of particles of ultrathin sections, of scanning electron microscopy, and related techniques. To quote the authors, "'studying electron microscope techniques in any text is not a substitute for actually performing the procedures in a laboratory with one's own hands". The reviewer believes this explicitly. Electron microscopy is best learned by apprenticeship. There are few principals that can be learned by reading a book, but these principles, such as they are, are all to be found in the article by Breese and Zacharia. The final chapter, "'Negative Staining for Electron Microscopy" is by DePamphilis. This again on Formvar coated grids, carbon film, and negative staining. Negative staining may have its place, but in this reviewer's experience. it is the cause of an unusual number of artifacts. 91
92
Book Re~iews
For those cases in which artit:acts are believed to not be induced by negative staining. DePamphilis's chapter is to be recommended. Overall. this is an excellent volume: full of information. lots of literature cited, lots of actually how to do-it-your-
self. and all in a nice small ~olume. which o n b costs 24.75 dollars. Dicision of Biology JAMES BONNER California Institute of Technology Pasadena. CA 91 i09. U.S.A.
Membrane Mediated Information--1. Biochemical Functions--Edited b.v P. W. Kent. American Elsevier, New York. 1973. 231 pp. 515.50. "'What's in a name?" or. indeed in a title'? The ~olume revie~cd hcrc contains a series of papers linked b? the common theme of study of oligosaccharide chains present at the surface of biological lipid membranes. These polymers are considered both in their own right, and as components of more complex and larger polymers, glycoproteins and glycolipids, Animal cells, especially cultured mammalian cell lines receive the greatest attention from the authors, though a three chapter section on viruses offers a view of membrane components of some enveloped viruses. "Information" is discussed in the opening article, by the late Richard Winzler. who points the number of permutations possible in a structure as simple as a disaccharide (16. in contrast to only 2 for a dipeptide). Many of the other papers tend to enlarge or detail for us the repertoire of oligosaccharide structure, but on the whole the readers are left to their own resources and to make their own inferences as to the "'informational" or biological meaning of structural variations in short sugar chains. (Important exceptions to them are the chapter on heterophile agglutinins and ABH blood group antigens dealing with systems in which both chemistry and immunology is understood). This reviewer appreciated such freedom to speculate, or perhaps ponder, and believes that the volume is valuable
as a special sort of extended review, of great use to biologists of all sorts, who study' the cell surface in terms of function, or serological specificity (but often in ignorance of chemical details) and conversely to chemists in search of functions and roles for particle structures. Most of the chapters combine brief reviews of an area with descriptions of fairly recent experimental work. Such combinations allow rapid entry into an area, e.g. glycolipid structure in transformed and non-transformed cells, together with a good appreciation of the technical aspects of work, the details of experiments too often omitted from more strictly review essays. Thus. though many of the experiments described here have been published in more extended lbrm elsewhere, they serve to reinforce the review aspects of the volume. Membrane glycolipids and glycoproteins, and some aspects of membrane phospholipids are dealt with in compact and adequate fashion. I would recommend the volume as what might be termed a "'working introduction" to an aspect of cell surface chemistry which, I predict will intrude itself more and more into cell biology.
Biology Department The Johns Hopkins Unicersirv Baltimore, MD 21218. U.S.A.
MICHAEL EDIDIN
Cellular Selection and Regulation in the Immune Response---Edited by Gerald M. Edelman. North Holland PuNishing Company, Amsterdam, 1974, 330 pp., S23.10. The collection of papers gathered together in this volume provide stimulating and interesting reading. A wide range of topics is covered in considerable detail there being 17 papers on the following themes: quantitation of clonal selection, immunogenetics, the cell surface and various aspects of lymphocyte stimulation, immunoglobulin synthesis and regulation of the immune response. It would be unfair to pick out articles as being good since the majority of articles are of a uniformly high standard and are a good combination of fact and speculation. One of the papers was. however, conspicuous in the absence of sufficient statistical qualification of the data presented in its numerous tables. Most of the information being presented in each article is not now new. but several of the papers each constitute a short review of their specified areas. They would therefore, provide a ready reference source for someone who quickly wants to be brought up to date with several different aspects of immunology. A few examples should give some idea of the scope covered in each section. Jerne in his paper 'Clonal Selection in a Lymphocyte Network' outlines the difficulties of quantitating clonal selection and discusses the possibilities of building a biologically acceptable mathematical model of the response of the immune response to antigens. Edelman in the same section considers the origin and mechanism of specificity in clonal selection b.~ covering
possible evolutionary relationships between histocompatibility loci. flz microglobulin and immunoglobulin; antibody specificity: the requirements of clonal selection and the ontogeny of specific antibody binding cells. The immunogenetics section includes articles on both histocompatibility linked genetic control of the immune response (McDevitt et al.) and the genetic fine structure of the H-2 complex (Shreffler). The former paper reviews and discusses evidence for and against the hypothesis that H-linked Ir genes are expressed only on T derived antigen reactive ceils. The latter paper contains a useful section for those unfamiliar with this field on the construction of the genetic map of the H-2 region of the, chromosome and describes the production, characteristics and potential uses of antisera to antigens controlled by genes localised in several papers but particular b that of Milstein et ,1., Histocompatibility antigens and other surface markers are used as tools to probe membrane structure. The binding of Ag-Ab complexes to cells and the possible role of this as a controlling function in the immune response is discussed. The proliferation and maturation of lymphocytes, the turnover of surface Ig, its site of synthesis and its relationship to secreted Ig are also covered in some depth. The various characteristics of Con A, its capacity to be both a stimulator and an inhibitor, its detailed structure