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

J. Biol. Chem., Vol. 276, Issue 10, 7357-7365, March 9, 2001

This Article Full Text Full Text (PDF) All Versions of this Article: 276/10/7357    most recent M009131200v1 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 Donnet, C. Articles by Sweadner, K. J. Search for Related Content PubMed PubMed Citation Articles by Donnet, C. Articles by Sweadner, K. J. Social Bookmarking                      What's this?

Thermal Denaturation of the Na,K-ATPase Provides Evidence for - Oligomeric Interaction and  Subunit Association with the C-terminal Domain^*

Claudia Donnet, Elena Arystarkhova, and Kathleen J. Sweadner

From the Laboratory of Membrane Biology, Neuroscience Center, Massachusetts General Hospital, Charlestown, Massachusetts 02129

Thermal denaturation can help elucidate protein domain substructure. We previously showed that the Na,K-ATPase partially unfolded when heated to 55 °C (Arystarkhova, E., Gibbons, D. L., and Sweadner, K. J. (1995) J. Biol. Chem. 270, 8785-8796). The  subunit unfolded without leaving the membrane, but three transmembrane spans (M8-M10) and the C terminus of the  subunit were extruded, while the rest of  retained its normal topology with respect to the lipid bilayer. Here we investigated thermal denaturation further, with several salient results. First, trypsin sensitivity at both surfaces of  was increased, but not sensitivity to V8 protease, suggesting that the cytoplasmic domains and extruded domain were less tightly packed but still retained secondary structure. Second, thermal denaturation was accompanied by SDS-resistant aggregation of  subunits as dimers, trimers, and tetramers without  or  subunits. This implies specific - contact. Third, the  subunit, like the C-terminal spans of , was selectively lost from the membrane. This suggests its association with M8-M10 rather than the more firmly anchored transmembrane spans. The picture that emerges is of a Na,K-ATPase complex of , , and  subunits in which  can associate in assemblies as large as tetramers via its cytoplasmic domain, while  and  subunits associate with  primarily in its C-terminal portion, which has a unique structure and thermal instability.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				D. Pavlovic, W. Fuller, and M. J. Shattock The intracellular region of FXYD1 is sufficient to regulate cardiac Na/K ATPase FASEB J, May 1, 2007; 21(7): 1539 - 1546. [Abstract] [Full Text] [PDF]

				A. Zouzoulas and R. Blostein Regions of the Catalytic {alpha} Subunit of Na,K-ATPase Important for Functional Interactions with FXYD 2 J. Biol. Chem., March 31, 2006; 281(13): 8539 - 8544. [Abstract] [Full Text] [PDF]

				M. Lindzen, K.-E. Gottschalk, M. Fuzesi, H. Garty, and S. J. D. Karlish Structural Interactions between FXYD Proteins and Na+,K+-ATPase: {alpha}/beta/FXYD SUBUNIT STOICHIOMETRY AND CROSS-LINKING J. Biol. Chem., March 3, 2006; 281(9): 5947 - 5955. [Abstract] [Full Text] [PDF]

				K. Geering FXYD proteins: new regulators of Na-K-ATPase Am J Physiol Renal Physiol, February 1, 2006; 290(2): F241 - F250. [Abstract] [Full Text] [PDF]

				K. Abe, S. Kaya, K. Taniguchi, Y. Hayashi, T. Imagawa, M. Kikumoto, K. Oiwa, and K. Sakaguchi Evidence for a Relationship between Activity and the Tetraprotomeric Assembly of Solubilized Pig Gastric H/K-ATPase J. Biochem., September 1, 2005; 138(3): 293 - 301. [Abstract] [Full Text] [PDF]

				D. H. Jones, T. Y. Li, E. Arystarkhova, K. J. Barr, R. K. Wetzel, J. Peng, K. Markham, K. J. Sweadner, G.-H. Fong, and G. M. Kidder Na,K-ATPase from Mice Lacking the {gamma} Subunit (FXYD2) Exhibits Altered Na+ Affinity and Decreased Thermal Stability J. Biol. Chem., May 13, 2005; 280(19): 19003 - 19011. [Abstract] [Full Text] [PDF]

				M. Fuzesi, K.-E. Gottschalk, M. Lindzen, A. Shainskaya, B. Kuster, H. Garty, and S. J. D. Karlish Covalent Cross-links between the {gamma} Subunit (FXYD2) and {alpha} and {beta} Subunits of Na,K-ATPase: MODELING THE {alpha}-{gamma} INTERACTION J. Biol. Chem., May 6, 2005; 280(18): 18291 - 18301. [Abstract] [Full Text] [PDF]

				C. Li, A. Grosdidier, G. Crambert, J.-D. Horisberger, O. Michielin, and K. Geering Structural and Functional Interaction Sites between Na,K-ATPase and FXYD Proteins J. Biol. Chem., September 10, 2004; 279(37): 38895 - 38902. [Abstract] [Full Text] [PDF]

				M. Laughery, M. Todd, and J. H. Kaplan Oligomerization of the Na,K-ATPase in Cell Membranes J. Biol. Chem., August 27, 2004; 279(35): 36339 - 36348. [Abstract] [Full Text] [PDF]

				A. V. Ivanov, M. E. Gable, and A. Askari Interaction of SDS with Na+/K+-ATPase: SDS-SOLUBILIZED ENZYME RETAINS PARTIAL STRUCTURE AND FUNCTION J. Biol. Chem., July 9, 2004; 279(28): 29832 - 29840. [Abstract] [Full Text] [PDF]

				Y. A. Mahmmoud, G. Cramb, A. B Maunsbach, C. P. Cutler, L. Meischke, and F. Cornelius Regulation of Na,K-ATPase by PLMS, the Phospholemman-like Protein from Shark: MOLECULAR CLONING, SEQUENCE, EXPRESSION, CELLULAR DISTRIBUTION, AND FUNCTIONAL EFFECTS OF PLMS J. Biol. Chem., September 26, 2003; 278(39): 37427 - 37438. [Abstract] [Full Text] [PDF]

				C. J. Costa, C. Gatto, and J. H. Kaplan Interactions between Na,K-ATPase alpha -Subunit ATP-binding Domains J. Biol. Chem., March 7, 2003; 278(11): 9176 - 9184. [Abstract] [Full Text] [PDF]

				M. J. Palladino, J. E. Bower, R. Kreber, and B. Ganetzky Neural Dysfunction and Neurodegeneration in Drosophila Na+/K+ ATPase Alpha Subunit Mutants J. Neurosci., February 15, 2003; 23(4): 1276 - 1286. [Abstract] [Full Text] [PDF]

				G. Crambert and K. Geering FXYD Proteins: New Tissue-Specific Regulators of the Ubiquitous Na,K-ATPase Sci. Signal., January 21, 2003; 2003(166): re1 - re1. [Abstract] [Full Text] [PDF]

				S. Teramachi, T. Imagawa, S. Kaya, and K. Taniguchi Replacement of Several Single Amino Acid Side Chains Exposed to the Inside of the ATP-binding Pocket Induces Different Extents of Affinity Change in the High and Low Affinity ATP-binding Sites of Rat Na/K-ATPase J. Biol. Chem., September 27, 2002; 277(40): 37394 - 37400. [Abstract] [Full Text] [PDF]

				U. Hasler, G. Crambert, J.-D. Horisberger, and K. Geering Structural and Functional Features of the Transmembrane Domain of the Na,K-ATPase beta Subunit Revealed by Tryptophan Scanning J. Biol. Chem., May 4, 2001; 276(19): 16356 - 16364. [Abstract] [Full Text] [PDF]