title: The transmembrane domain of glycoprotein Ibbeta is critical to efficient expression of glycoprotein Ib-IX complex in the plasma membrane.
abstract: Lack of expression of glycoprotein (GP) Ib-IX-V complex in platelets often results from mutations in its three subunits: GP Ibalpha, GP Ibbeta, or GP IX. The requirement of all three subunits in the efficient surface expression of the receptor complex has been reproduced in Chinese hamster ovary cells. Here, we probed the role of the transmembrane domains in expression of the GP Ib-IX complex and potential interactions between these domains. Replacing the transmembrane domains of either GP Ibalpha or GP Ibbeta, but not that of GP IX, with unrelated sequences markedly diminished surface expression of the GP Ib-IX complex in transiently transfected Chinese hamster ovary cells. Replacement of the Ibbeta transmembrane domain produced the largest effect. Furthermore, several single-site mutations in the Ibbeta transmembrane domain were found to significantly decrease overall expression as well as surface expression of GP Ibalpha, probably by perturbing the interaction between the Ibalpha and Ibbeta transmembrane domains and in turn reducing the stability of GP Ibalpha in the cell. Mutations S503V and S503L in the Ibalpha transmembrane domain partly reversed the expression-decreasing effect of mutation H139L, but not the others, in the Ibbeta transmembrane domain, suggesting a specific interaction between these two polar residues. Together, our results have demonstrated the importance of the Ibbeta transmembrane domain, through its interaction with the Ibalpha counterpart, to the proper assembly and efficient surface expression of the GP Ib-IX complex.
authors.affiliation: Center for Membrane Biology, Department of Biochemistry and Molecular Biology, University of Texas Medical School, Houston, Texas 77030, USA.
authors.forename: Xi
authors.initials: X
authors.lastname: Mo
authors.forename: Nan
authors.initials: N
authors.lastname: Lu
authors.forename: Arnoldo
authors.initials: A
authors.lastname: Padilla
authors.forename: José A
authors.initials: JA
authors.lastname: López
authors.forename: Renhao
authors.initials: R
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism*; D006224:Cricetinae; D005786:Gene Expression Regulation; D006639:Histidine / Q000737:chemistry; D008969:Molecular Sequence Data; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D012694:Serine / Q000737:chemistry
chemical_list: D019038:Platelet Glycoprotein GPIb-IX Complex; D012694:Serine; D006639:Histidine