Structural, thermodynamic, and phosphatidylinositol 3-phosphate binding properties of Phafin2

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dc.contributor.authorTang, TuoXianen
dc.contributor.authorJo, Amien
dc.contributor.authorDeng, Jingrenen
dc.contributor.authorEllena, Jeffrey F.en
dc.contributor.authorLazar, Iuliana M.en
dc.contributor.authorDavis, Richey M.en
dc.contributor.authorCapelluto, Daniel G. S.en
dc.date.accessioned2017-06-13T12:34:31Zen
dc.date.available2017-06-13T12:34:31Zen
dc.date.issued2017-04-01en
dc.description.abstractPhafin2 is a phosphatidylinositol 3-phosphate (PtdIns(3)P) binding protein involved in the regulation of endosomal cargo trafficking and lysosomal induction of autophagy. Binding of Phafin2 to PtdIns(3)P is mediated by both its PH and FYVE domains. However, there are no studies on the structural basis, conformational stability, and lipid interactions of Phafin2 to better understand how this protein participates in signaling at the surface of endomembrane compartments. Here, we show that human Phafin2 is a moderately elongated monomer of ~28 kDa with an intensity-average hydrodynamic diameter of ~7 nm. Circular dichroism (CD) analysis indicates that Phafin2 exhibits an a/b structure and predicts ~40% random coil content in the protein. Heteronuclear NMR data indicates that a unique conformation of Phafin2 is present in solution and dispersion of resonances suggests that the protein exhibits random coiled regions, in agreement with the CD data. Phafin2 is stable, displaying a melting temperature of 48.48C. The folding-unfolding curves, obtained using urea- and guanidine hydrochloride-mediated denaturation, indicate that Phafin2 undergoes a two-state native-to-denatured transition. Analysis of these transitions shows that the free energy change for urea- and guanidine hydrochloride-induced Phafin2 denaturation in water is ~4 kcal mol21. PtdIns(3)P binding to Phafin2 occurs with high affinity, triggering minor conformational changes in the protein. Taken together, these studies represent a platform for establishing the structural basis of Phafin2 molecular interactions and the role of the two potentially redundant PtdIns(3)P-binding domains of the protein in endomembrane compartments.en
dc.description.versionPublished versionen
dc.format.extent814 - 823 (10) page(s)en
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.1002/pro.3128en
dc.identifier.issn0961-8368en
dc.identifier.issue4en
dc.identifier.orcidDavis, RM [0000-0002-4838-2541]en
dc.identifier.orcidCapelluto, DGS [0000-0002-0412-4508]en
dc.identifier.urihttp://hdl.handle.net/10919/78032en
dc.identifier.volume26en
dc.languageEnglishen
dc.publisherWileyen
dc.relation.urihttp://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000398183800015&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1en
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectBiochemistryen
dc.subjectPhafin2en
dc.subjectphosphatidylinositol 3-phosphateen
dc.subjectconformationen
dc.subjectprotein structureen
dc.subjectSEDIMENTATION-VELOCITY EXPERIMENTSen
dc.subjectFYVE DOMAINen
dc.subjectMolecular Biologyen
dc.subjectAPOPTOSISen
dc.titleStructural, thermodynamic, and phosphatidylinositol 3-phosphate binding properties of Phafin2en
dc.title.serialProtein Scienceen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
pubs.organisational-group/Virginia Techen
pubs.organisational-group/Virginia Tech/All T&R Facultyen
pubs.organisational-group/Virginia Tech/Engineeringen
pubs.organisational-group/Virginia Tech/Engineering/Chemical Engineeringen
pubs.organisational-group/Virginia Tech/Engineering/COE T&R Facultyen
pubs.organisational-group/Virginia Tech/Faculty of Health Sciencesen
pubs.organisational-group/Virginia Tech/Scienceen
pubs.organisational-group/Virginia Tech/Science/Biological Sciencesen
pubs.organisational-group/Virginia Tech/Science/COS T&R Facultyen
pubs.organisational-group/Virginia Tech/University Research Institutesen
pubs.organisational-group/Virginia Tech/University Research Institutes/Fralin Life Sciencesen
pubs.organisational-group/Virginia Tech/University Research Institutes/Fralin Life Sciences/Fralin Affiliated Facultyen

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