HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Uchida, E. Articles by Anraku, Y. Search for Related Content PubMed PubMed Citation Articles by Uchida, E. Articles by Anraku, Y. Social Bookmarking                      What's this?

J. Biol. Chem., Vol. 263, Issue 1, 45-51, Jan, 1988

Characterization and function of catalytic subunit alpha of H+- translocating adenosine triphosphatase from vacuolar membranes of Saccharomyces cerevisiae. A study with 7-chloro-4-nitrobenzo-2-oxa-1,3- diazole

E Uchida, Y Ohsumi and Y Anraku
Department of Biology, Faculty of Science, University of Tokyo, Japan.

Subunit alpha (Mr 89,000) from vacuolar membrane H+-translocating adenosine triphosphatase of the yeast Saccharomyces cerevisiae was found to bind 8-azido[alpha-32P]adenosine triphosphate. Labeling by this photosensitive ATP derivative was saturable with an apparent dissociation constant of 10(-6) to 10(-5) M and decreased in the presence of ATP and ADP. The enzyme was inactivated by 7-chloro-4- nitrobenzo-2-oxa-1,3-diazole (NBD-Cl), with about 1 microM causing half- maximal inactivation in the neutral pH range. This inactivation was prevented by the presence of ATP, ADP, or adenosyl-5'-yl imidodiphosphate (AMP-PNP). The original activity was restored by treating the inactivated enzyme with 2-mercaptoethanol. Kinetic and chemical studies of the inactivation showed that the activity was lost on chemical modification of a single tyrosine residue per molecule of the enzyme. When the enzyme was inactivated with [14C]NBD-Cl, subunit alpha was specifically labeled, and this labeling was completely prevented by the presence of ATP, GTP, ADP, or AMP-PNP. From these results, it was concluded that subunit alpha of yeast vacuolar H+- ATPase has a catalytic site that contains a single, essential tyrosine residue. The kinetics of single site hydrolysis of [gamma-32P]ATP (Grubmeyer, C., Cross, R. L., and Penefsky, H. S. (1982) J. Biol. Chem. 257, 12092-12100) indicated the formation of an enzyme-ATP complex and subsequent hydrolysis of bound ATP to ADP and Pi at the NBD-Cl- sensitive catalytic site. NBD-Cl inactivated the single site hydrolysis and inhibited the formation of an enzyme-ATP complex. Dicyclohexylcarbodiimide did not affect the single site hydrolysis, but inhibited the enzyme activity under steady-state conditions.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				T. Hirata, N. Nakamura, H. Omote, Y. Wada, and M. Futai Regulation and Reversibility of Vacuolar H+-ATPase J. Biol. Chem., January 7, 2000; 275(1): 386 - 389. [Abstract] [Full Text] [PDF]

				M Futai, T Oka, G Sun-Wada, Y Moriyama, H Kanazawa, and Y Wada Luminal acidification of diverse organelles by V-ATPase in animal cells J. Exp. Biol., January 1, 2000; 203(1): 107 - 116. [Abstract]

				C. Forster, M. A. Santos, S. Ruffert, R. Kramer, and J. L. Revuelta Physiological Consequence of Disruption of the VMA1 Gene in the Riboflavin Overproducer Ashbya gossypii J. Biol. Chem., April 2, 1999; 274(14): 9442 - 9448. [Abstract] [Full Text] [PDF]

				K. J. Parra and P. M. Kane Reversible Association between the V1 and V0 Domains of Yeast Vacuolar H+-ATPase Is an Unconventional Glucose-Induced Effect Mol. Cell. Biol., December 1, 1998; 18(12): 7064 - 7074. [Abstract] [Full Text]

				E. Vasilyeva and M. Forgac Interaction of the Clathrin-coated Vesicle V-ATPase with ADP and Sodium Azide J. Biol. Chem., September 11, 1998; 273(37): 23823 - 23829. [Abstract] [Full Text] [PDF]

				R. Hirata, L. A. Graham, A. Takatsuki, T. H. Stevens, and Y. Anraku VMA11 and VMA16 Encode Second and Third Proteolipid Subunits of the Saccharomyces cerevisiae Vacuolar Membrane H+-ATPase J. Biol. Chem., February 21, 1997; 272(8): 4795 - 4803. [Abstract] [Full Text] [PDF]

				K. J. Parra and P. M. Kane Wild-type and Mutant Vacuolar Membranes Support pH-dependent Reassembly of the Yeast Vacuolar H+-ATPase in Vitro J. Biol. Chem., August 9, 1996; 271(32): 19592 - 19598. [Abstract] [Full Text] [PDF]

				E. Vasilyeva and M. Forgac 3'-O-(4-Benzoyl)benzoyladenosine 5'-Triphosphate Inhibits Activity of the Vacuolar (H+)-ATPase from Bovine Brain Clathrin-coated Vesicles by Modification of a Rapidly Exchangeable, Noncatalytic Nucleotide Binding Site on the B Subunit J. Biol. Chem., May 31, 1996; 271(22): 12775 - 12782. [Abstract] [Full Text] [PDF]

				J. J. Tomashek, J. L. Sonnenburg, J. M. Artimovich, and D. J. Klionsky Resolution of Subunit Interactions and Cytoplasmic Subcomplexes of the Yeast Vacuolar Proton-translocating ATPase J. Biol. Chem., April 26, 1996; 271(17): 10397 - 10404. [Abstract] [Full Text] [PDF]

				C. Landolt-Marticorena, K. M. Williams, J. Correa, W. Chen, and M. F. Manolson Evidence That the NH2 Terminus of Vph1p, an Integral Subunit of the V0 Sector of the Yeast V-ATPase, Interacts Directly with the Vma1p and Vma13p Subunits of the V1 Sector J. Biol. Chem., May 12, 2000; 275(20): 15449 - 15457. [Abstract] [Full Text] [PDF]