Vacuolar H+-pyrophosphatase (H+-PPase) from etiolated hypocotyls of mung bean (Vigna radiata L.) is a homodimer with a molecular mass of 145 kDa. The vacuolar H+-PPase was subjected to high hydrostatic pressure to investigate its structure and function. The inhibition of H+-PPase activity by high hydrostatic pressure has a pressure-, time- and protein-concentration-dependent manner. The Vmax value of vacuolar H+-PPase was dramatically decreased by pressurization from 293.9 to 70.2 µmol of PPi (pyrophosphate) consumed/h per mg of protein, while the Km value decreased from 0.35 to 0.08 mM, implying that the pressure treatment increased the affinity of PPi to vacuolar H+-PPase but decreased its hydrolysis. The physiological substrate and its analogues enhance high pressure inhibition of vacuolar H+-PPase. The HPLC profile reveals high pressure treatment of H+-PPase provokes the subunit dissociation from an active into inactive form. High hydrostatic pressure also induces the conformational change of vacuolar H+-PPase as determined by spectroscopic techniques. Our results indicate the importance of protein–protein interaction for this novel proton-translocating enzyme. Working models are proposed to interpret the pressure inactivation of vacuolar H+-PPase. We also suggest that association of identical subunits of vacuolar H+-PPase is not random but proceeds in a specific manner.
Skip Nav Destination
Article navigation
April 1998
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
Research Article|
April 15 1998
Subunit interaction of vacuolar H+-pyrophosphatase as determined by high hydrostatic pressure
Su J. YANG;
Su J. YANG
1Institute of Radiation Biology, College of Nuclear Science, National Tsing Hua University, Hsin Chu 30043, Taiwan, Republic of China
Search for other works by this author on:
Shu J. KO;
Shu J. KO
1Institute of Radiation Biology, College of Nuclear Science, National Tsing Hua University, Hsin Chu 30043, Taiwan, Republic of China
Search for other works by this author on:
Yuan R. TSAI;
Yuan R. TSAI
1
1Institute of Radiation Biology, College of Nuclear Science, National Tsing Hua University, Hsin Chu 30043, Taiwan, Republic of China
Search for other works by this author on:
Shih S. JIANG;
Shih S. JIANG
1Institute of Radiation Biology, College of Nuclear Science, National Tsing Hua University, Hsin Chu 30043, Taiwan, Republic of China
Search for other works by this author on:
Soong Y. KUO;
Soong Y. KUO
1Institute of Radiation Biology, College of Nuclear Science, National Tsing Hua University, Hsin Chu 30043, Taiwan, Republic of China
Search for other works by this author on:
Shu H. HUNG;
Shu H. HUNG
1Institute of Radiation Biology, College of Nuclear Science, National Tsing Hua University, Hsin Chu 30043, Taiwan, Republic of China
Search for other works by this author on:
Rong L. PAN
Rong L. PAN
2
1Institute of Radiation Biology, College of Nuclear Science, National Tsing Hua University, Hsin Chu 30043, Taiwan, Republic of China
2To whom correspondence should be addressed (e-mail rlpan@RB.nthu.edu.tw).
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
June 30 1997
Revision Received:
November 03 1997
Accepted:
January 15 1998
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1998
1998
Biochem J (1998) 331 (2): 395–402.
Article history
Received:
June 30 1997
Revision Received:
November 03 1997
Accepted:
January 15 1998
Citation
Su J. YANG, Shu J. KO, Yuan R. TSAI, Shih S. JIANG, Soong Y. KUO, Shu H. HUNG, Rong L. PAN; Subunit interaction of vacuolar H+-pyrophosphatase as determined by high hydrostatic pressure. Biochem J 15 April 1998; 331 (2): 395–402. doi: https://doi.org/10.1042/bj3310395
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Could not validate captcha. Please try again.