A new membrane-associated Ca2+-binding protein of rat spermatogenic cells

Its purification and characterization

Masahisa Nakamura, Tomoyo Yamanobe, Takashi Suyemitsu, Masayuki Komukai, Ryuichi Kan, Shoichi Okinaga, Kiyoshi Arai

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

A Ca2+-binding protein of Mr=52000, estimated by SDS-PAGE, was purified to a final yield of 0.04% from rat spermatogenic cells. Purification steps included gel filtration, ammonium sulfate precipitation and HPLC. Amino acid analysis showed the content of 34% acidic residues and 15% basic residues. The isoelectric point of this protein was 4.7. Dot-blot analysis indicated that the Ca2+-binding protein bound 2 mol of calcium per mol of protein. This protein had two binding sites with dissociation constants of 4.8 μM and 0.2 μM. No appreciable amount of hexose was observed (<1 μg of hexose/70 ug of protein). This protein may play an important role such as the Ca2+-transport in the plasma membrane of spermatogenic cells.

Original languageEnglish
Pages (from-to)1358-1364
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume176
Issue number3
DOIs
Publication statusPublished - 1991 May 15
Externally publishedYes

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Purification
Rats
Carrier Proteins
Membranes
Hexoses
Proteins
Isoelectric Point
Ammonium Sulfate
Cell membranes
Gel Chromatography
Polyacrylamide Gel Electrophoresis
Gels
Binding Sites
High Pressure Liquid Chromatography
Cell Membrane
Calcium
Amino Acids

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

A new membrane-associated Ca2+-binding protein of rat spermatogenic cells : Its purification and characterization. / Nakamura, Masahisa; Yamanobe, Tomoyo; Suyemitsu, Takashi; Komukai, Masayuki; Kan, Ryuichi; Okinaga, Shoichi; Arai, Kiyoshi.

In: Biochemical and Biophysical Research Communications, Vol. 176, No. 3, 15.05.1991, p. 1358-1364.

Research output: Contribution to journalArticle

Nakamura, Masahisa ; Yamanobe, Tomoyo ; Suyemitsu, Takashi ; Komukai, Masayuki ; Kan, Ryuichi ; Okinaga, Shoichi ; Arai, Kiyoshi. / A new membrane-associated Ca2+-binding protein of rat spermatogenic cells : Its purification and characterization. In: Biochemical and Biophysical Research Communications. 1991 ; Vol. 176, No. 3. pp. 1358-1364.
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