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

J. Biol. Chem., Vol. 281, Issue 16, 10760-10768, April 21, 2006

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: 281/16/10760    most recent M513783200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted 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 Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Takahashi, K. Articles by Ueda, K. Search for Related Content PubMed PubMed Citation Articles by Takahashi, K. Articles by Ueda, K. Social Bookmarking                      What's this?

Purification and ATPase Activity of Human ABCA1^*

From the Laboratory of Cellular Biochemistry, Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan

ATP-binding cassette protein A1 (ABCA1) plays a major role in cholesterol homeostasis and high density lipoprotein metabolism. Apolipoprotein A-I binds to ABCA1 and cellular cholesterol and phospholipids, mainly phosphatidylcholine, are loaded onto apoA-I to form pre- high density lipoprotein (HDL). It is proposed that ABCA1 translocates phospholipids and cholesterol directly or indirectly to form pre- HDL. To explore the mechanism of ABCA1-mediated pre- HDL formation, we expressed human ABCA1 in insect Sf9 cells and purified it. Trypsin limited-digestion of purified ABCA1 in the detergent-soluble form suggested that it retained conformation similar to ABCA1 expressed in the membranes of human fibroblast WI-38 cells. Purified ABCA1 showed robust ATPase activity when reconstituted in liposomes made of synthetic phosphatidylcholine. ABCA1 showed lower ATPase activity when reconstituted in liposomes containing phosphatidylserine, phosphatidylethanolamine, or phosphatidylglycerol and also showed weak specificity in acyl chain species. ATPase activity was reduced by the addition of cholesterol and decreased by 25% in the presence of 20% cholesterol. -Sitosterol and campesterol showed similar inhibitory effects but stigmasterol did not, suggesting structure-specific interaction between ABCA1 and sterols. Glibenclamide suppressed ABCA1 ATPase, suggesting that it inhibits apoA-I-dependent cellular cholesterol efflux by suppressing ABCA1 ATPase activity. These results suggest that the ATPase activity of ABCA1 is stimulated preferentially by phospholipids with choline head groups, phosphatidylcholine and sphingomyelin. This study with purified human ABCA1 provides the first biochemical basis of the mechanism for HDL formation mediated by ABCA1.

Received for publication, December 27, 2005 , and in revised form, February 15, 2006.

^* This work was supported by Grant-in-aid for Scientific Research and Creative Scientific Research 15GS0301 from the Ministry of Education, Culture, Sports, Science, and Technology, Japan, and grants from the Bio-oriented Technology Research Advancement Institution (BRAIN), and the Pharmaceutical and Medical Devices Agency. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. 1– 4.

¹ To whom correspondence should be addressed. Tel.: 81-75-753-6124; Fax: 81-75-753-6104; E-mail: uedak{at}kais.kyoto-u.ac.jp.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				C. Radojkovic, A. Genoux, V. Pons, G. Combes, H. de Jonge, E. Champagne, C. Rolland, B. Perret, X. Collet, F. Terce, et al. Stimulation of Cell Surface F1-ATPase Activity by Apolipoprotein A-I Inhibits Endothelial Cell Apoptosis and Promotes Proliferation Arterioscler. Thromb. Vasc. Biol., July 1, 2009; 29(7): 1125 - 1130. [Abstract] [Full Text] [PDF]

				K. Nagao, Y. Zhao, K. Takahashi, Y. Kimura, and K. Ueda Sodium taurocholate-dependent lipid efflux by ABCA1: effects of W590S mutation on lipid translocation and apolipoprotein A-I dissociation J. Lipid Res., June 1, 2009; 50(6): 1165 - 1172. [Abstract] [Full Text] [PDF]

				M. Hozoji, Y. Kimura, N. Kioka, and K. Ueda Formation of Two Intramolecular Disulfide Bonds Is Necessary for ApoA-I-dependent Cholesterol Efflux Mediated by ABCA1 J. Biol. Chem., April 24, 2009; 284(17): 11293 - 11300. [Abstract] [Full Text] [PDF]

				A. R. Tanaka, F. Kano, A. Yamamoto, K. Ueda, and M. Murata Formation of cholesterol-enriched structures by aberrant intracellular accumulation of ATP-binding cassette transporter A1. Genes Cells, August 1, 2008; 13(8): 889 - 904. [Abstract] [Full Text] [PDF]

				K. Nagao, K. Takahashi, K. Hanada, N. Kioka, M. Matsuo, and K. Ueda Enhanced ApoA-I-dependent Cholesterol Efflux by ABCA1 from Sphingomyelin-deficient Chinese Hamster Ovary Cells J. Biol. Chem., May 18, 2007; 282(20): 14868 - 14874. [Abstract] [Full Text] [PDF]

				J. Wang, F. Sun, D.-w. Zhang, Y. Ma, F. Xu, J. D. Belani, J. C. Cohen, H. H. Hobbs, and X.-S. Xie Sterol Transfer by ABCG5 and ABCG8: IN VITRO ASSAY AND RECONSTITUTION J. Biol. Chem., September 22, 2006; 281(38): 27894 - 27904. [Abstract] [Full Text] [PDF]

				D. Trompier, M. Alibert, S. Davanture, Y. Hamon, M. Pierres, and G. Chimini Transition from Dimers to Higher Oligomeric Forms Occurs during the ATPase Cycle of the ABCA1 Transporter J. Biol. Chem., July 21, 2006; 281(29): 20283 - 20290. [Abstract] [Full Text] [PDF]