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Transcript
Membrane Protein Structure and Assembly Ross Dalbey The Ohio State University Department of Chemistry Structures and architectures • • • • • Protein-lipid interactions Two architectures (at least) TM helices Connecting loops and tails The “positive inside” rule Biological Membrane Molecular Interactions in Membrane Proteins Heijne Nature Reviews Molecular Cell Biology 7, 909–918 (December 2006) | doi:10.1038/nrm2063 Only two basic structures TM helix lengths are typically 20-30 residues (Bowie, JMB 272: 780) Loops tend to be short (Tusnady & Simon, JMB 283:488) The “positive inside” rule C N Periplasm Inner Membrane N C Cytoplasm C N Procoat Leader Peptidase Dalbey (1990) TIBS 15, 253-257 MalF Topology can be manipulated C N Periplasm Inner Membrane N N Cytoplasm Assembly of Membrane Proteins Cyto III+IV I II Cytochrome bo oxidase LacY permease F1Fo ATPase Many membranes in a eukaryotic cell Membrane translocation and insertion devices in bacteria and in the endoplasmic reticulum The machinery that is involved in membrane protein insertion in E. coli Xie and Dalbey (2008) Nature Reviews Micro 6, 235 The X-ray Structure of the SecY complex in Methanococcus jannaschii Front to Front Van den berg et al., 2004, Nature 427 36-44 Van den Berg et al (2004) Nature 427, 36 Different stages of translocation of a secretory protein Recognition of a transmembrane helix by the Sec61 translocon Hessa et al (2005) Nature 433, 377 Evolutionarily conserved YidC Family Members Stuart and Neupert (2000) Nature (News and Views) 406, 575 Jim Samuelson YidC is essential for cell viability Samuelson et al., (2000) Nature, 406, 637-41 YidC depletion studies Samuelson et al., 2000, Nature, 406, 637-641 Effect of YidC depletion on Secindependent Membrane Insertion Samuelson et al., 2000, Nature, 406, 637-641 Illustration of site-specific photocrosslinking YidC and Ffh interact with Pf3 coat Chen et al. (2002) JBC, 277, 7670-7675 YidC is a Membrane Protein Insertase for Sec-Independent Proteins YidC - + N PMF Procoat Insertion N- - + + C Pf3 coat Insertion Dalbey and Kuhn (2004) J. Cell Biol. 166, 769 + + C Why is YidC essential for Bacteria? H+ H+ H+ H+ Cytochrome NADHOxidase dehydroUbiquinol bo3 genases Pool ee e- NADH NAD+ H+ O2 Periplasm F0 Cytoplasm H2O F1 ATP synthase Respiratory chain complexes in E. coli Substrates for the YidC-only pathway N N N Pf3coat M13procoat Periplasm N F-ATPase subunit c MscL •YidC is required for membrane insertion of these substrates •Pf3 coat can insert into proteoliposomes containing only YidC Substrates for the YidC-Sec pathway P1 C N Periplasm C N CyoA F1Fo-ATPase subunit a C N LacY •YidC is required for membrane insertion of CyoA and subunit a •YidC is required for the membrane protein folding of LacY YidC functions in the integration of TS into the membrane Xie and Dalbey (2008) Nature Reviews Micro 6, 235 YidC functions in the folding of Lac permease Nagamori et al. (2004) J. Cell Biol. 165, 53 Assembly of cytochrome bo oxidase Dalbey et al (2011) Ann Rev Biochem, in press. Assembly of F1Fo ATP synthase Dalbey et al (2011) Ann Rev Biochem, in press Conclusion • α-helices and β-sheets are the basic membrane-spanning regions • Positively charged residues control the membrane protein topology • Sec translocase and YidC insertase are the devices used to insert proteins • The bacterial YidC functions as a membrane chaperone and assembly factor