Protamines. Isolation, Characterization, Structure and Function.
Authors: T Ando, M Yamasaki, K Suzuki
Journal: Annals of Internal Medicine (1974)

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  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.