Molecular cloning of a rat liver cDNA encoding the 16 kDa subunit of vacuolar H+-ATPases: Organellar and tissue distribution of 16 kDa proteolipids

Jun Ichi Nezu, Kiyoto Motojima, Hiro Omi Tamura, Shoji Ohkuma

研究成果: Article査読

30 被引用数 (Scopus)

抄録

A cDNA (T3-L) encoding the 16 kDa subunit of vacuolar H+-ATPase was cloned from a cDNA library of rat liver. A polypeptide of 156 amino acids with a molecular mass of 15,807 Da (pI=9.5) having four hydrophobic stretches was predicted. T3-L polypeptide was 92% and 100% identical with the 16 kDa proteolipid of bovine chromaffin granule and that of mouse, respectively. Antisera raised against the NH2-terminal of the T3-L polypeptide reacted positively with the membrane ghosts of rat liver tritosomes and the partially purified H+-ATPase thereof. Western blotting of subcellular fractions with the antisera showed high abundance of 16 kDa protein in the lysosomes, although a significant amount was also detected in the Golgi apparatus. Western blotting of rat tissues revealed high levels of 16 kDa proteolipid in the brain and the kidney. Northern blots with T3-L similarly showed considerably high expression of T3-L mRNA in the brain and the kidney. Southern hybridization of rat genomic DNA with T3-L showed at most three distinct bands, regardless of the stringency of hybridization and whether hybridization was performed with its subfragments. This suggests the possibility of multiple (at least three) homologous/identical genes encoding 16 kDa proteolipid. The possible presence and significance of isoforms of 16 kDa proteolipid in rats are discussed.

本文言語English
ページ(範囲)212-219
ページ数8
ジャーナルJournal of biochemistry
112
2
DOI
出版ステータスPublished - 1992 8月
外部発表はい

ASJC Scopus subject areas

  • 生化学
  • 分子生物学

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