title: Membrane topography and topogenesis of prenylated Rab acceptor (PRA1).
abstract: The mouse prenylated Rab acceptor (mPRA1) is associated with the Golgi membrane at steady state and interacts with Rab proteins. It contains two internal hydrophobic domains (34 residues each) that have enough residues to form four transmembrane (TM) segments. In this study, we have determined the membrane topography of mPRA1 in both intact cells and isolated microsomes. The putative TM segments of mPRA1 were used to substitute for a known TM segment of a model membrane protein to determine whether the mPRA1 segments integrate into the membrane. Furthermore, N-linked glycosylation scanning methods were used to distinguish luminal domains from cytoplasmic domains of mPRA1. The data demonstrate that mPRA1 is a polytopic membrane protein containing four TM segments. These TM segments act cooperatively during the translocation and integration at the endoplasmic reticulum membrane. All hydrophilic domains are in the cytoplasm, including the N-terminal domain, the linker domain between the two hydrophobic domains, and the C-terminal domain. As a result, the bulk of mPRA1 is located in the cytoplasm, supporting its postulated role in regulating Rab membrane targeting and intracellular trafficking.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA. jialing-lin@ouhsc.edu
authors.forename: J
authors.initials: J
authors.lastname: Lin
authors.forename: Z
authors.initials: Z
authors.lastname: Liang
authors.forename: Z
authors.initials: Z
authors.lastname: Zhang
authors.forename: G
authors.initials: G
authors.lastname: Li
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry*; D002462:Cell Membrane / Q000737:chemistry*; D003593:Cytoplasm / Q000737:chemistry; D006031:Glycosylation; D051379:Mice; D011817:Rabbits; D033921:Vesicular Transport Proteins
chemical_list: D002352:Carrier Proteins; C498435:Rabac1 protein, mouse; D033921:Vesicular Transport Proteins