title: Cleavage of a multispanning membrane protein by an intramembrane serine protease.
abstract: All intramembrane proteases are known to cleave membrane proteins with a single transmembrane helix. Such cleavages often release anchored soluble domains, which play a role in physiologically important inter- and intracellular processes. However, in many cases the physiological roles/substrates of intramembrane proteases are not known. It is interesting that no multispanning substrates were identified so far, despite the fact that intramembrane proteases have promiscuous substrate recognition and cleavage capabilities. Here we determined whether, in a synthetic experimental system, intramembrane proteases have the capability to interact with and cleave multispanning membrane proteins. We utilized the Escherichia coli rhomboid GlpG, an intramembrane serine protease, and truncated versions of the E. coli multidrug transporter MdfA as model multispanning membrane proteins. On the basis of in vivo and in vitro studies on the association of GlpG with various MdfA constructs and their cleavage, we conclude that GlpG is able to recognize and cleave truncated forms of MdfA but not the intact protein. We propose that GlpG has the capacity to act on unfolded multispanning membrane proteins, thus providing an incentive for investigating possible physiological consequences.
authors.affiliation: Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot 76100, Israel.
authors.forename: Elinor
authors.initials: E
authors.lastname: Erez
authors.forename: Eitan
authors.initials: E
authors.lastname: Bibi
mesh_terms: D000595:Amino Acid Sequence; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010450:Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D017433:Protein Structure, Secondary
chemical_list: D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C102898:GlpG protein, E coli; C118438:Mdfa protein, E coli; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D010450:Endopeptidases