Boar transition protein 2 and 4 isolated from late spermatid nuclei by high-performance liquid chromatography.
The boar late spermatid nuclei retaining transition proteins (TPs) could be obtained from the testis by the use of antipain to inhibit TP-degrading proteinases of the nuclei. The enzymes detected in acid extract including the basic proteins were inactivated by reduction and carboxymethylation of the proteins. The reduced and carboxymethylated basic proteins were fractionated by differential precipitation between 3% trichloroacetic acid (TCA) and 3-20% TCA. From the 3% TCA-precipitate, boar TP2 and TP4 were isolated by high-performance liquid chromatography (HPLC) on Nucleosil 300 7C18. The two TPs were characterized by acid urea- and SDS-polyacrylamide gel electrophoreses and amino acid analysis. Boar TP2 closely resembled rat and mouse TP2s, and ram protein 3 in its high content of serine and basic amino acids, the presence of cysteine and molecular weight. Boar TP4 was similar to ram protein P1 in its high content of basic amino acids, the presence of cysteine and molecular weight. But the TP2 and TP4 differed in electrophoretic mobility on acid urea-gel and solubility in 3% TCA from those of the other species. The HPLC used here also enabled us to efficiently separate boar TP1, TP2, TP3 and TP4, and to estimate that the amount of the TP2, TP3 and TP4 was about 1/8, 1/4 and 1/4 that of the TP1, respectively.
Akama
K
K
Department of Chemistry, Faculty of Science, Chiba University, Japan.
Maruyama
R
R
Mochizuki
H
H
Tobita
T
T
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002467:Cell Nucleus / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D008297:Male; D008970:Molecular Weight; D013045:Species Specificity; D013087:Spermatids / Q000737:chemistry*; D013552:Swine
D000596:Amino Acids; D002868:Chromosomal Proteins, Non-Histone; C053886:spermatid transition proteins