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Conference Paper: Solution structures and dynamics of the sterile α Motif (SAM) domain of the deleted in liver cancer 2 (DLC2): a monomeric structure with membrane-binding properties

TitleSolution structures and dynamics of the sterile α Motif (SAM) domain of the deleted in liver cancer 2 (DLC2): a monomeric structure with membrane-binding properties
Authors
Issue Date2005
Citation
Croucher Foundation Advanced Study Institute: Advances in Protein Sciences, Biochemistry Department, The Chinese University of Hong Kong, Hong Kong, 15-17 December, p. 33, abstract no. PO-17 How to Cite?
AbstractThe Deleted in Liver Cancer 2 (DLC2) gene encodes a Rho GTPase-activating protein (GAP) with growth suppressor function. In addition to the RhoGAP domain, this molecule also contains a sterile α motif (SAM) and a lipid-binding StAR-related lipid-transfer (START) domain. To gain an insight into the function of DLC2, we have expressed and purified a recombinant 13C/15N doubly-labeled DLC2 SAM domain. The three-dimensional solution structure and dynamics of the SAM domain of DLC2, DLC2-SAM, were investigated by NMR spectroscopy together with molecular dynamic simulated annealing. We showed that DLC2-SAM is distinct from all other known SAM domains by the presence of a four-helix bundle. The spatial arrangement of four α-helices is also unique. Backbone dynamics studies showed a restricted motion of four helices and a high mobility of the N-and C-termini as well as loops. Although it has been shown that some members of the SAM domain family can form dimers and oligomers, both NMR and biochemical analyses indicated that DLC2-SAM exists as a monomer in solution. We also examined the interaction of DLC2-SAM domain with sodium dodecyl sulfate (SDS) micelles by NMR and CD spectroscopic techniques. 2D [1H,15N] HSQC spectra of DLC2-SAM in the presence of SDS displayed shifts of most residues although still well dispersed, in agreement with slightly decreased α-helical content based on CD spectra of DLC2-SAM in the absence and presence of SDS, suggesting that DLC2-SAM may interact with membrane lipids in vivo with secondary structure changes of the protein. Finally, we demonstrated that DLC2-SAM is dispensable for the self association of full-length DLC2 in vivo.
Persistent Identifierhttp://hdl.handle.net/10722/95963

 

DC FieldValueLanguage
dc.contributor.authorLi, Hen_HK
dc.contributor.authorFung, KLen_HK
dc.contributor.authorJin, Den_HK
dc.contributor.authorChung, SSMen_HK
dc.contributor.authorCing, YPen_HK
dc.contributor.authorNg, IOLen_HK
dc.contributor.authorSze, KHen_HK
dc.contributor.authorKo, CBen_HK
dc.contributor.authorSun, Hen_HK
dc.date.accessioned2010-09-25T16:19:02Z-
dc.date.available2010-09-25T16:19:02Z-
dc.date.issued2005en_HK
dc.identifier.citationCroucher Foundation Advanced Study Institute: Advances in Protein Sciences, Biochemistry Department, The Chinese University of Hong Kong, Hong Kong, 15-17 December, p. 33, abstract no. PO-17en_HK
dc.identifier.urihttp://hdl.handle.net/10722/95963-
dc.description.abstractThe Deleted in Liver Cancer 2 (DLC2) gene encodes a Rho GTPase-activating protein (GAP) with growth suppressor function. In addition to the RhoGAP domain, this molecule also contains a sterile α motif (SAM) and a lipid-binding StAR-related lipid-transfer (START) domain. To gain an insight into the function of DLC2, we have expressed and purified a recombinant 13C/15N doubly-labeled DLC2 SAM domain. The three-dimensional solution structure and dynamics of the SAM domain of DLC2, DLC2-SAM, were investigated by NMR spectroscopy together with molecular dynamic simulated annealing. We showed that DLC2-SAM is distinct from all other known SAM domains by the presence of a four-helix bundle. The spatial arrangement of four α-helices is also unique. Backbone dynamics studies showed a restricted motion of four helices and a high mobility of the N-and C-termini as well as loops. Although it has been shown that some members of the SAM domain family can form dimers and oligomers, both NMR and biochemical analyses indicated that DLC2-SAM exists as a monomer in solution. We also examined the interaction of DLC2-SAM domain with sodium dodecyl sulfate (SDS) micelles by NMR and CD spectroscopic techniques. 2D [1H,15N] HSQC spectra of DLC2-SAM in the presence of SDS displayed shifts of most residues although still well dispersed, in agreement with slightly decreased α-helical content based on CD spectra of DLC2-SAM in the absence and presence of SDS, suggesting that DLC2-SAM may interact with membrane lipids in vivo with secondary structure changes of the protein. Finally, we demonstrated that DLC2-SAM is dispensable for the self association of full-length DLC2 in vivo.-
dc.languageengen_HK
dc.relation.ispartofCroucher Foundation Advanced Study Institute: Advances in Protein Sciences, Biochemistry Department, The Chinese University of Hong Kong, Hong Kong, 15-17 Decemberen_HK
dc.titleSolution structures and dynamics of the sterile α Motif (SAM) domain of the deleted in liver cancer 2 (DLC2): a monomeric structure with membrane-binding propertiesen_HK
dc.typeConference_Paperen_HK
dc.identifier.emailLi, H: hylichem@hkucc.hku.hken_HK
dc.identifier.emailJin, D: dyjin@hkucc.hku.hken_HK
dc.identifier.emailNg, IOL: iolng@hkucc.hku.hken_HK
dc.identifier.emailSze, KH: khsze@hku.hken_HK
dc.identifier.emailKo, CB: cbko@hkucc.hku.hken_HK
dc.identifier.emailSun, H: hsun@hku.hken_HK
dc.identifier.authorityJin, D=rp00452en_HK
dc.identifier.authorityNg, IOL=rp00335en_HK
dc.identifier.authoritySze, KH=rp00785en_HK
dc.identifier.authoritySun, H=rp00777en_HK
dc.identifier.hkuros122689en_HK
dc.identifier.spage33-
dc.identifier.epage33en_HK

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