title: Fine resolution of human sperm nucleoproteins by two-dimensional electrophoresis.
abstract: Human sperm nucleoproteins consist of protamines and histones. Changes in composition of these proteins are thought to correlate with spermatogenesis and may be involved in some instances of male infertility. We sought to separate sperm nucleoproteins including variants of protamine using an improved two-dimensional electrophoretic method, with the aim of comprehensively analysing all sperm nucleoprotein constituents. After extracting nuclear basic proteins from the sperm of normal volunteers, we analysed these proteins on a gel sheet by a radical free, highly reducing method based on Kaltschmidt and Whittmann's two-dimensional electrophoresis. Basic proteins from sperm nuclei were separated clearly into 12 spots. By amino acid sequence analysis, these spots corresponded to protamine 1 (P1)- (five spots), protamine 2 (P2)-related proteins (six spots) and testis-specific histone H2B (one spot). The N-terminal amino acid sequences of the six P2-related proteins were compatible with those of HPI1, HPI2, HPS1, HPS2, HP2 and HP3, and quantitative comparison could be performed. In conclusion, human sperm nucleoproteins including all P2-related variants could be analysed quantitatively with high resolution on a single electrophoretic gel.
authors.affiliation: Department of Obstetrics and Gynecology, Keio University School of Medicine, Tokyo, Japan.
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Yoshii
authors.forename: Naoaki
authors.initials: N
authors.lastname: Kuji
authors.forename: Setsuko
authors.initials: S
authors.lastname: Komatsu
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Iwahashi
authors.forename: Yudai
authors.initials: Y
authors.lastname: Tanaka
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Yoshida
authors.forename: Akira
authors.initials: A
authors.lastname: Wada
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Yoshimura
mesh_terms: D000595:Amino Acid Sequence; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006657:Histones / Q000032:analysis / Q000737:chemistry; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009698:Nucleoproteins / Q000032:analysis / Q000737:chemistry*; D011479:Protamines / Q000032:analysis / Q000737:chemistry; D013094:Spermatozoa / Q000737:chemistry*
chemical_list: D006657:Histones; D009698:Nucleoproteins; C082073:PRM1 protein, human; D011479:Protamines; C068621:protamine 2