H+-transloaction ATP in Golgi apparatus. Characterization as vacuolar H+-ATPase and its subunit structures

Y. Moriyama, N. Nelson

Research output: Contribution to journalArticle

108 Citations (Scopus)

Abstract

Golgi apparatus was prepared from rat liver, and enzymatic properties and the subunit structure of the H+ATPase were characterized. GTP (and also ITP) was found to drive H+-transport with about 20% of the initial velocity as that of ATP. Bafilomycin, a specific inhibitor for vacuolar H+-ATPase, inhibited the activity at 2.5 nM. The H+-ATPase was completely inhibited in the cold in the presence of MgATP (5 mM) and NaNO3 (0.1 M). The cold inactivation of the H+-ATPase resulted in release of a set of polypeptides from Golgi membrane, with molecular masses almost identical to that of the hydrophilic sector of chromaffin granule H+-ATPase (72, 57, 41, 34, and 33 kDa). Three of these polypeptides (72, 57, and 34 kDa), cross-reacted with antibodies against the corresponding subunits of the chromaffin granule H+-ATPase. A counterpart of the 39-kDa hydrophobic component of chromaffin granule H+-ATPase was identified in the membrane, but no 115-kDa component was found. Hence, the Golgi H+-ATPase, shows typical features of vacuolar H+-ATPase, in relatively low substrate specificity, its repsonse to inhibitors, inactivation by cold treatment in the presence of MgATP, and subunit composition judged by antibody cross-reactivity.

Original languageEnglish
Pages (from-to)18445-18450
Number of pages6
JournalJournal of Biological Chemistry
Volume264
Issue number31
Publication statusPublished - 1989
Externally publishedYes

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Vacuolar Proton-Translocating ATPases
Proton-Translocating ATPases
Golgi Apparatus
Adenosine Triphosphate
Chromaffin Granules
Inosine Triphosphate
Membranes
Peptides
Antibodies
Molecular mass
Substrate Specificity
Guanosine Triphosphate
Liver
Rats
Substrates
Chemical analysis

ASJC Scopus subject areas

  • Biochemistry

Cite this

H+-transloaction ATP in Golgi apparatus. Characterization as vacuolar H+-ATPase and its subunit structures. / Moriyama, Y.; Nelson, N.

In: Journal of Biological Chemistry, Vol. 264, No. 31, 1989, p. 18445-18450.

Research output: Contribution to journalArticle

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