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Advances in cyclic nucleotide research, 10, current methodology
Cell, Vol. 18, 229-231,
September
1979,
Copyright
0 1979
by Ml7
Book Reviews
Expanding
Cyclic Nucleotides
Advances in Cyclic Nucleotide Research, 10, Current Methodology. G. Brooker, P. Greengard and G. A. Robison, eds. New York: Raven Press. 259 pp. $24.00.
Research on cyclic nucleotides has expanded to such an extent that it now touches upon a large variety of biological topics ranging from gene expression in bacteria to growth and differentiation in higher organisms, and the number of investigators engaged in cyclic nucleotide research has increased accordingly. It is therefore essential that a detailed description and evaluation of methodology for cyclic nucleotide research be available. Volume 10 in the series of Advances in Cyclic Nucleotide Research fulfills this need. This timely book is organized and written in such a manner that both beginners and experienced investigators in cyclic nucleotide research will find it of value and interest. Principles of assays and detailed descriptions of assay procedures are provided for the methods of analysis. Chapters are included which deal with the quantitation of cyclic AMP and cyclic GMP by radioimmunoassay, and the assay systems %or adenylate cyclase, guanylate cyclase and cyclic nucleotide phosphodiesterase. Cyclic nucleotide-dependent protein kinase assay procedures are described and an additional chapter detailing methods currently in use to evaluate endogenous phosphorylation in cell-free and intact cell preparations is presented. Additional methodology is provided for immunocytochemical localization of cyclic nucleotides and protein kinase and also for isolation of cyclic nucleotide receptor protein by affinity chromatography. A recent exciting advance related to cyclic nucleotide research is the discovery and evaluation of the Ca++ -dependent modulator protein. One chapter is devoted to the preparation and assay of this protein. It is well known that enzymes in cyclic nucleotide metabolism are activated by divalent cations and that substrate and product nucleotides act as chelating agents. A description of chelation theory and tabulations of chelation properties of nucleotides and the commonly used chelating agents, EDTA and EGTA, are presented. Tables in this chapter are useful for experiments in which it is essential to know the concentrations of bound and free cations. An informative chapter describing recent techniques for enzymatic preparation of labeled nucleotides with high specific activity is included as well. Those investigators on a limited research budget will find this chapter quite useful.
This recent volume in this popular for those studying cyclic nucleotide function.
series is a must metabolism and
George S. Johnson Laboratory of Molecular Biology National Cancer Institute Bethesda, Maryland 20205
Inhibiting
Protein Synthesis
Inhibitors of Protein Biosynthesis. Molecular Biology, Biochemistry and Biophysics, 30. By D. Vazquez. Berlin, Heidelberg, New York: Springer-Verlag. 312 pp. $32.50.
It has been said that if we could lay hands on specific inhibitors of all the metabolic processes we know, the task of elucidating those we do not understand would be that much easier. Pie in the sky, perhaps, but there can be few readers of this journal who have failed to use antibiotics or related substances as tools to inhibit some aspect of macromolecular biosynthesis at one time or another: mitomycin or FUDR to block DNA synthesis; actinomycin to inhibit gene expression by interfering with the template function of DNA; rifampicin to block transcription in procaryotic cells, or (Yamanitin to prevent transcription by eucaryotic RNA polymerase II; chloramphenicol or cycloheximide to inhibit translation of mRNA by procaryotic or eucaryotic cytoplasmic ribosomes, respectively. The list is long, and still longer if one includes such multipurpose agents as the classic intercalative DNA binder ethidium bromide (originally developed as an effective trypanocidal drug for veterinary use, remember, although now consumed on the kilogram scale in laboratories around the world for anything from isolating circular DNA to whacking mitochondria or staining gels). Specificity (or better, biochemical selectivity) is the crucial element in this story-not so much selective toxicity toward different cell types as originally envisaged by Ehrlich, although that is not without relevance to many an area of current research (to say nothing of human need as the strategy of chemotherapy moves into its fifth decade), but the capacity to discriminate effectively between biochemical processes. If necessary, conditions must be judiciously chosen so that the investigator can pinpoint the lesion he has introduced into the biological system in order to interpret
September
1979,
Copyright
0 1979
by Ml7
Book Reviews
Expanding
Cyclic Nucleotides
Advances in Cyclic Nucleotide Research, 10, Current Methodology. G. Brooker, P. Greengard and G. A. Robison, eds. New York: Raven Press. 259 pp. $24.00.
Research on cyclic nucleotides has expanded to such an extent that it now touches upon a large variety of biological topics ranging from gene expression in bacteria to growth and differentiation in higher organisms, and the number of investigators engaged in cyclic nucleotide research has increased accordingly. It is therefore essential that a detailed description and evaluation of methodology for cyclic nucleotide research be available. Volume 10 in the series of Advances in Cyclic Nucleotide Research fulfills this need. This timely book is organized and written in such a manner that both beginners and experienced investigators in cyclic nucleotide research will find it of value and interest. Principles of assays and detailed descriptions of assay procedures are provided for the methods of analysis. Chapters are included which deal with the quantitation of cyclic AMP and cyclic GMP by radioimmunoassay, and the assay systems %or adenylate cyclase, guanylate cyclase and cyclic nucleotide phosphodiesterase. Cyclic nucleotide-dependent protein kinase assay procedures are described and an additional chapter detailing methods currently in use to evaluate endogenous phosphorylation in cell-free and intact cell preparations is presented. Additional methodology is provided for immunocytochemical localization of cyclic nucleotides and protein kinase and also for isolation of cyclic nucleotide receptor protein by affinity chromatography. A recent exciting advance related to cyclic nucleotide research is the discovery and evaluation of the Ca++ -dependent modulator protein. One chapter is devoted to the preparation and assay of this protein. It is well known that enzymes in cyclic nucleotide metabolism are activated by divalent cations and that substrate and product nucleotides act as chelating agents. A description of chelation theory and tabulations of chelation properties of nucleotides and the commonly used chelating agents, EDTA and EGTA, are presented. Tables in this chapter are useful for experiments in which it is essential to know the concentrations of bound and free cations. An informative chapter describing recent techniques for enzymatic preparation of labeled nucleotides with high specific activity is included as well. Those investigators on a limited research budget will find this chapter quite useful.
This recent volume in this popular for those studying cyclic nucleotide function.
series is a must metabolism and
George S. Johnson Laboratory of Molecular Biology National Cancer Institute Bethesda, Maryland 20205
Inhibiting
Protein Synthesis
Inhibitors of Protein Biosynthesis. Molecular Biology, Biochemistry and Biophysics, 30. By D. Vazquez. Berlin, Heidelberg, New York: Springer-Verlag. 312 pp. $32.50.
It has been said that if we could lay hands on specific inhibitors of all the metabolic processes we know, the task of elucidating those we do not understand would be that much easier. Pie in the sky, perhaps, but there can be few readers of this journal who have failed to use antibiotics or related substances as tools to inhibit some aspect of macromolecular biosynthesis at one time or another: mitomycin or FUDR to block DNA synthesis; actinomycin to inhibit gene expression by interfering with the template function of DNA; rifampicin to block transcription in procaryotic cells, or (Yamanitin to prevent transcription by eucaryotic RNA polymerase II; chloramphenicol or cycloheximide to inhibit translation of mRNA by procaryotic or eucaryotic cytoplasmic ribosomes, respectively. The list is long, and still longer if one includes such multipurpose agents as the classic intercalative DNA binder ethidium bromide (originally developed as an effective trypanocidal drug for veterinary use, remember, although now consumed on the kilogram scale in laboratories around the world for anything from isolating circular DNA to whacking mitochondria or staining gels). Specificity (or better, biochemical selectivity) is the crucial element in this story-not so much selective toxicity toward different cell types as originally envisaged by Ehrlich, although that is not without relevance to many an area of current research (to say nothing of human need as the strategy of chemotherapy moves into its fifth decade), but the capacity to discriminate effectively between biochemical processes. If necessary, conditions must be judiciously chosen so that the investigator can pinpoint the lesion he has introduced into the biological system in order to interpret