Protamines

A century has already passed since FRIEDRICH MIESCHER, working at Strasbourg and Basel, began his study of protamine, one of the basic nuclear proteins of cells. It was first established by KOSSEL that protamine represents the simplest known protein. In the conviction that research into the nature of protamine would shed light on that of other typical proteins, a group of researchers in Germany followed MIESCHER and laid the foundations of protein chemistry. A general view of prot­ amines was thus built up by KOSSEL, working at Strasbourg, Berlin, Marburg an der Lahn, and Heidelberg, FELIX at Heidelberg, Munich, and Frankfurt am Main, and WALDSCHMIDT-LEITZ at Prague and Munich. Concepts and techniques established by these studies have been widely utilized for research on other typical proteins. The revolutionary advances in chemical and physical techniques after W orId War II extended the sphere of research to Tokyo in the Far East. Prof. FELIX' visit in 1955 greatly encouraged our research group in Tokyo. His death in August 1960 constituted a sad loss to protein chemistry and stimulated our group to assume responsibility for carrying on the studies. In the following decade we in Tokyo have been able to add a new development to the results on the chemical structure of protamines accumulated by the Eurqpean researchers over a period of about fifty years.

Contenu

I Introduction.- (Accompanied by a Select Bibliography on Protamines, Histones, and Nucleoproteins).- II Distribution of Nucleoprotamines and Protamines.- III Preparation of Nucleoprotamines and Protamines.- A. Isolation of Sperm Heads.- 1. Outline of the "Classical" Procedure of Kossel.- 2. Isolation of Sperm Heads by Fractional Centrifugation in Isotonic Salt Solution.- 3. "Plasmolysis" Procedure of Felix.- 4. Isolation of Sperm Heads and/or Cell Nucleus Fraction or Chromatin from Fish Testes during Spermatogenesis.- B. Isolation and Purification of Nucleoprotamine.- C. Separation and Purification of Protamines from Sperm Heads or Nucleoprotamine.- 1. Classical Procedures.- a) Extraction with Dilute Mineral Acid.- b) Extraction with Aqueous Cupric Salt.- c) Further Purification of Protamine Preparations.- 2. Modern Improvements to Classical Procedures.- 3. Some Remarks on the Methods of Preparation of Whole Protamines.- IV Composition.- A. Composition of Nucleoprotamines.- 1. Composition of Sperm Head or Sperm Nucleus.- 2. Composition of Reconstituted Nucleoprotamine.- 3. Composition of Artificial Nucleoprotamines.- 4. Base Composition of Deoxyribonucleic Acid Associated with Protamine.- B. Composition of Unfractionated (or Whole) Protamines.- 1. Classification of Protamines.- 2. Ratio of Carbon to Nitrogen in Protamine.- 3. Amino-Acid Composition.- V Molecular Weight.- A. Nucleoprotamines.- B. Protamines (Whole or Unfractionated).- VI Chemical Structure of Nucleoprotamines and Protamines.- A. Nucleoprotamines.- B. Protamines (Whole or Unfractionated).- 1. Amino-Terminal Residue and Sequence.- 2. Carboxyl-Terminal Residue and Sequence.- 3. Amino-Acid Sequences of Peptides Obtained by Partial Hydrolysis - Hypothetical Structures of Whole Protamines.- VII Heterogeneity of Protamines and Homogeneous Molecular Species of Protamines.- A. Heterogeneity in Protamines.- B. Preparative Fractionation of Protamines into Their Components.- 1. Fractionation of Whole Clupeine into Y and Z Fractions by Column Chromatography on Buffered Alumina (Homogeneous Clupeine Z).- 2. Fractionation of Whole Clupeine into Y and Z Fractions by Column Chromatography on CM-Cellulose (Homogeneous Clupeine Z).- 3. Fractionation of Trinitrophenylated (TNP-) Whole Clupeine into TNP-YI, TNP-Z and Free YII Components by Column Chromatography on CM-Cellulose (Homogeneous TNP-Clupeine YI, TNP-Clupeine Z and Free Clupeine YII).- 4. Fractionation of Whole Clupeine into Its Three Components, YI, YII and Z, and of Whole Salmine and Iridine into Some of Their Components by One-Step Elution Chromatography on a Column of CM-Sephadex or Bio-Gel CM (Homogeneous Clupeine YI, YII and Z; Homogeneous Salmine AI, Iridine I and II).- 5. Fractionation of Whole Thynnine into Four Fractions, Y1, Y2, Zl and Z2, by Chromatography on a Column of CM-Sephadex (Homogeneous Thynnine Y1, Y2, Zl and Z2).- 6. Fractionation of Whole Galline into Several Components by Column Chromatography on Bio-Gel CM-30.- 7. Isolation and Partial Characterization of a Basic Protein from Bull Sperm Heads.- VIII Chemical Structure of Homogeneous Molecular Species of Protamines.- A. Determination of the Complete Amino-Acid Sequence of Clupeine Z.- 1. Amino-Acid Composition, Amino Terminus, and Molecular Weight.- 2. Analysis of Tryptic Peptides of Clupeine Z, Leading to a Partial Chemical Structure.- 3. Application of N?O Acyl Rearrangement Reaction to Clupeine Z, Followed by Selective Chemical Cleavage of the Chain, Leading to the Final Primary Structure.- B. The Amino-Acid Sequence of Clupeine YII.- C. The Amino-Acid Sequence of Clupeine YI.- D. Comments on the Structure of Clupeine.- E. The Amino-Acid Sequences of the Three Components (Y'I, Y?II and Z?) of Clupeine from North Sea Herring.- F. The Amino-Acid Sequences of One Component of Salmine and Three Components of Iridine.- IX Physical Structure of Nucleoprotamines and Protamines.- A. Nucleoprotamines.- B. Protamines.- X Properties and Functions.- A. Physical Properties.- B. Chemical Properties.- C. Biological and Physiological Properties.- 1. Biological Functions.- 2. Physiological Properties.- Acknowledgement.- References.