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 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 Hsu, H. H. Search for Related Content PubMed PubMed Citation Articles by Hsu, H. H. Social Bookmarking                      What's this?

J. Biol. Chem., Vol. 258, Issue 6, 3463-3468, Mar, 1983

Purification and partial characterization of ATP pyrophosphohydrolase from fetal bovine epiphyseal cartilage

HH Hsu

ATP pyrophosphohydrolase was partially purified from fetal bovine epiphyseal cartilage. The purification was about 10- and 100-fold over the enzyme activities of matrix vesicle fraction and cell homogenate, respectively. The pyrophosphohydrolase and alkaline phosphatase were separated by a sequential application of Sepharose CL-6B and DEAE- cellulose column chromatographies. The purified enzyme migrated as a single band corresponding to the molecular weight of 230,000 in sodium dodecyl sulfate-polyacrylamide disc gel by electrophoresis. The enzyme absolutely required Zn2+ for its activity and appeared to bind Zn2+ strongly with an apparent affinity of p[Zn2+]0.5 = 13.4. The apparent Km for ATP was 0.18 mM. The enzyme was also reactive toward various nucleoside triphosphates including GTP, CTP, and UTP. In contrast, various phosphodiesters including RNA, UDP-glucose, NAD, and bis-p- nitrophenylphosphate were 5% or less as reactive as the nucleoside triphosphates. The pyrophosphohydrolase was inactive toward adenosine 3':5'-monophosphate or various phosphonates. UDP-glucose (1 mM), NAD (1 mM), or RNA (1 mg/ml) failed to inhibit the ATP pyrophosphohydrolase activity. These observations suggest that the ATP pyrophosphohydrolase of the cartilage is probably not a phosphodiesterase I. The matrix vesicle fraction, which probably also included some plasma membrane vesiculated during collagenase digestion, contained the highest specific activity of the enzyme as compared to other subcellular fractions of either epiphyseal or articular cartilage.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M. M. McCormick, F. Rahimi, Y. V. Bobryshev, K. Gaus, H. Zreiqat, H. Cai, R. S. A. Lord, and C. L. Geczy S100A8 and S100A9 in Human Arterial Wall: IMPLICATIONS FOR ATHEROGENESIS J. Biol. Chem., December 16, 2005; 280(50): 41521 - 41529. [Abstract] [Full Text] [PDF]

				L. Zhang, M. Balcerzak, J. Radisson, C. Thouverey, S. Pikula, G. Azzar, and R. Buchet Phosphodiesterase Activity of Alkaline Phosphatase in ATP-initiated Ca2+ and Phosphate Deposition in Isolated Chicken Matrix Vesicles J. Biol. Chem., November 4, 2005; 280(44): 37289 - 37296. [Abstract] [Full Text] [PDF]

				H. C. Anderson, J. B. Sipe, L. Hessle, R. Dhamyamraju, E. Atti, N. P. Camacho, and J. L. Millan Impaired Calcification Around Matrix Vesicles of Growth Plate and Bone in Alkaline Phosphatase-Deficient Mice Am. J. Pathol., March 1, 2004; 164(3): 841 - 847. [Abstract] [Full Text] [PDF]

				H. H.T. Hsu and H. C. Anderson Evidence of the Presence of a Specific ATPase Responsible for ATP-initiated Calcification by Matrix Vesicles Isolated from Cartilage and Bone J. Biol. Chem., October 18, 1996; 271(42): 26383 - 26388. [Abstract] [Full Text] [PDF]