Kinetic studies of chromaffin granule H+-ATPase and effects of bafilomycin A1

Hironori Hanada, Yoshinori Moriyama, Masatomo Maeda, Masamitsu Futai

Research output: Contribution to journalArticle

106 Citations (Scopus)

Abstract

Vacuolar type H+-ATPase purified from bovine chromaffin granules did not show simple Michaelis-Menten type kinetics, and had apparent Km values of 5 μM, 30 μM and 300 μM. These three Km values suggested the presence of catalytic cooperativity during steady-state hydrolysis. The single turnover rate was 10-3 -fold the maximal velocity of the enzyme and similar to the rate estimated from the velocity of steady-state hydrolysis with the smallest Km value (5 μM). The H+-ATPase was inhibited by the stoichiometric binding of bafilomycin A1, a specific inhibitor of vacuolar type H+-ATPase. This inhibitor not only lowered the rate of ATP hydrolysis at the single catalytic site, but also affected the catalytic cooperativity of the enzyme.

Original languageEnglish
Pages (from-to)873-878
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume170
Issue number2
DOIs
Publication statusPublished - Jul 31 1990
Externally publishedYes

Fingerprint

Chromaffin Granules
Proton-Translocating ATPases
Vacuolar Proton-Translocating ATPases
Hydrolysis
Kinetics
Enzymes
Catalytic Domain
Adenosine Triphosphate
bafilomycin A1

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Kinetic studies of chromaffin granule H+-ATPase and effects of bafilomycin A1. / Hanada, Hironori; Moriyama, Yoshinori; Maeda, Masatomo; Futai, Masamitsu.

In: Biochemical and Biophysical Research Communications, Vol. 170, No. 2, 31.07.1990, p. 873-878.

Research output: Contribution to journalArticle

Hanada, Hironori ; Moriyama, Yoshinori ; Maeda, Masatomo ; Futai, Masamitsu. / Kinetic studies of chromaffin granule H+-ATPase and effects of bafilomycin A1. In: Biochemical and Biophysical Research Communications. 1990 ; Vol. 170, No. 2. pp. 873-878.
@article{775334906dc6429da5426017e717382f,
title = "Kinetic studies of chromaffin granule H+-ATPase and effects of bafilomycin A1",
abstract = "Vacuolar type H+-ATPase purified from bovine chromaffin granules did not show simple Michaelis-Menten type kinetics, and had apparent Km values of 5 μM, 30 μM and 300 μM. These three Km values suggested the presence of catalytic cooperativity during steady-state hydrolysis. The single turnover rate was 10-3 -fold the maximal velocity of the enzyme and similar to the rate estimated from the velocity of steady-state hydrolysis with the smallest Km value (5 μM). The H+-ATPase was inhibited by the stoichiometric binding of bafilomycin A1, a specific inhibitor of vacuolar type H+-ATPase. This inhibitor not only lowered the rate of ATP hydrolysis at the single catalytic site, but also affected the catalytic cooperativity of the enzyme.",
author = "Hironori Hanada and Yoshinori Moriyama and Masatomo Maeda and Masamitsu Futai",
year = "1990",
month = "7",
day = "31",
doi = "10.1016/0006-291X(90)92172-V",
language = "English",
volume = "170",
pages = "873--878",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Kinetic studies of chromaffin granule H+-ATPase and effects of bafilomycin A1

AU - Hanada, Hironori

AU - Moriyama, Yoshinori

AU - Maeda, Masatomo

AU - Futai, Masamitsu

PY - 1990/7/31

Y1 - 1990/7/31

N2 - Vacuolar type H+-ATPase purified from bovine chromaffin granules did not show simple Michaelis-Menten type kinetics, and had apparent Km values of 5 μM, 30 μM and 300 μM. These three Km values suggested the presence of catalytic cooperativity during steady-state hydrolysis. The single turnover rate was 10-3 -fold the maximal velocity of the enzyme and similar to the rate estimated from the velocity of steady-state hydrolysis with the smallest Km value (5 μM). The H+-ATPase was inhibited by the stoichiometric binding of bafilomycin A1, a specific inhibitor of vacuolar type H+-ATPase. This inhibitor not only lowered the rate of ATP hydrolysis at the single catalytic site, but also affected the catalytic cooperativity of the enzyme.

AB - Vacuolar type H+-ATPase purified from bovine chromaffin granules did not show simple Michaelis-Menten type kinetics, and had apparent Km values of 5 μM, 30 μM and 300 μM. These three Km values suggested the presence of catalytic cooperativity during steady-state hydrolysis. The single turnover rate was 10-3 -fold the maximal velocity of the enzyme and similar to the rate estimated from the velocity of steady-state hydrolysis with the smallest Km value (5 μM). The H+-ATPase was inhibited by the stoichiometric binding of bafilomycin A1, a specific inhibitor of vacuolar type H+-ATPase. This inhibitor not only lowered the rate of ATP hydrolysis at the single catalytic site, but also affected the catalytic cooperativity of the enzyme.

UR - http://www.scopus.com/inward/record.url?scp=0024991786&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024991786&partnerID=8YFLogxK

U2 - 10.1016/0006-291X(90)92172-V

DO - 10.1016/0006-291X(90)92172-V

M3 - Article

VL - 170

SP - 873

EP - 878

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 2

ER -