Multistate GTPase Control Co-translational Protein Targeting by Xin Zhang

By Xin Zhang

Proteins act as macromolecular equipment that mediate many varied organic procedures - the molecular mechanisms of this equipment has interested biologists for many years. research of the kinetic and thermodynamic gains of those mechanisms might display unheard of features of ways the equipment functionality and should finally bring about a singular knowing of varied organic techniques. This dissertation comprehensively demonstrates how universally conserved guanosine triphosphatases within the sign reputation particle and its membrane receptor keep the potency and constancy of the co-translational protein focusing on strategy necessary to all cells. a sequence of quantitative experiments demonstrate that the hugely ordered and coordinated conformational states of the equipment are the major to their regulatory functionality. This dissertation additionally bargains a mechanistic view of one other attention-grabbing approach within which multistate protein equipment heavily keep watch over severe organic processes.

Written whereas finishing graduate paintings at California Institute of Technology.

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DTT was removed by gel filtration chromatography. The reduced and degassed proteins (*100 lM) were labeled with a three- to fivefold molar excess of MTSSL (Toronto Research Chemicals, Toronto, Canada) at room temperature in the dark for 2–3 h. Excess MTSSL was removed by gel filtration chromatography. The labeling efficiency was determined by EPR using the TEMPO calibration curve (Bruker user manual), and was typically [80% with \5% background as assessed from the cysteine-less wildtype protein using the same procedure.

35. 36. 37. 38. 39. 40. 41. 42. 43. 44. 45. 46. 47. 48. Walter P, Johnson AE (1994) Annu Rev Cell Biol 10:87 Matlack KES, Mothes W, Rapoport TA (1998) Cell 92:381 Johnson AE, van Waes MA (1999) Annu Rev Cell Dev Biol 15:799 Shan SO, Walter P (2005) FEBS Lett 579:921 Shan SO, Stroud RM, Walter P (2004) PLoS Biol 2:1572 Shan SO, Chandrasekar S, Walter P (2007) J Cell Biol 178:611 Walter P, Ibrahimi I, Blobel G (1981) J Cell Biol 91:545 Walter P, Blobel G (1981) J Cell Biol 91:551 Walter P, Blobel G (1981) J Cell Biol 91:557 Batey RT, Rambo RP, Lucast L, Rha B, Doudna JA (2000) Science 287:1232 Keenan RJ, Freymann DM, Walter P, Stroud RM (1998) Cell 94:181 Krieg UC, Walter P, Johnson AE (1986) Proc Natl Acad Sci U S A 83:8604 Kurzchalia TV et al (1986) Nature 320:634 Zopf D, Bernstein HD, Johnson AE, Walter P (1990) EMBO J 9:4511 Connolly T, Rapiejko PJ, Gilmore R (1991) Science 252:1171 Egea PF et al (2004) Nature 427:215 Peluso P et al (2000) Science 288:1640 Powers T, Walter P (1995) Science 269:1422 Cleverley RM, Gierasch LM (2002) J Biol Chem 277:46763 Bourne HR, Sanders DA, Mccormick F (1991) Nature 349:117 Gawronski-Salerno J, Coon JSV, Focia PJ, Freymann DM (2007) Proteins 66:984 Focia PJ, Gawronski-Salerno J, Coon JS, Freymann DM (2006) J Mol Biol 360:631 Freymann DM, Keenan RJ, Stroud RM, Walter P (1999) Nat Struct Biol 6:793 Freymann DM, Keenan RJ, Stroud RM, Walter P (1997) Nature 385:361 Peluso P, Shan SO, Nock S, Herschlag D, Walter P (2001) Biochem 40:15224 Gu SQ et al (2005) RNA-Publ RNA Soc 11:1374 Siu FY, Spanggord RJ, Doudna JA (2007) RNA 13:240 Sagar MB, Lucast L, Doudna JA (2004) RNA 10:772 Brown S, Fournier MJ (1984) J Mol Biol 178:533 Jagath JR et al (2001) RNA-Publ RNA Soc 7:293 Zheng N, Gierasch LM (1997) Mol Cell 1:79 Poritz MA, Strub K, Walter P (1988) Cell 55:4 Poritz MA et al (1990) Science 250:1111 Batey RT, Sagar MB, Doudna JA (2001) J Mol Biol 307:229 Hainzl T, Huang S, Sauer-Eriksson AE (2007) Proc Natl Acad Sci U S A 104:14911 Stroud RM, Walter P (1999) Curr Opin Struct Biol 9:754 Spanggord RJ, Siu F, Ke AL, Doudna JA (2005) Nat Struct Mol Biol 12:1116 Mandon EC, Jiang Y, Gilmore R (2003) J Cell Biol 162:575 Buskiewicz I et al (2005) J Mol Biol 351:417 Powers T, Walter P (1997) EMBO J 16:4880 Connolly T, Gilmore R (1989) Cell 57:599 Rapiejko PJ, Gilmore R (1992) J Cell Biol 117:493 Zopf D, Bernstein HD, Walter P (1993) J Cell Biol 120:1113 Jagath JR, Rodnina MV, Wintermeyer W (2000) J Mol Biol 295:745 Padmanabhan S, Freymann DM (2001) Structure 9:859 Ramirez UD et al (2002) J Mol Biol 320:783 Focia PJ, Alam H, Lu T, Ramirez UD, Freymann DM (2004) Proteins 54:222 Lakowicz JR (1999) Principles of fluorescence spectroscopy.

14. 15. 16. 17. 18. 19. 20. 21. 22. 23. 24. 25. 26. 27. 28. 29. 30. 31. 32. 33. 34. 35. 36. 37. 38. 39. 40. 41. 42. 43. 44. 45. 46. 47. 48. Walter P, Johnson AE (1994) Annu Rev Cell Biol 10:87 Matlack KES, Mothes W, Rapoport TA (1998) Cell 92:381 Johnson AE, van Waes MA (1999) Annu Rev Cell Dev Biol 15:799 Shan SO, Walter P (2005) FEBS Lett 579:921 Shan SO, Stroud RM, Walter P (2004) PLoS Biol 2:1572 Shan SO, Chandrasekar S, Walter P (2007) J Cell Biol 178:611 Walter P, Ibrahimi I, Blobel G (1981) J Cell Biol 91:545 Walter P, Blobel G (1981) J Cell Biol 91:551 Walter P, Blobel G (1981) J Cell Biol 91:557 Batey RT, Rambo RP, Lucast L, Rha B, Doudna JA (2000) Science 287:1232 Keenan RJ, Freymann DM, Walter P, Stroud RM (1998) Cell 94:181 Krieg UC, Walter P, Johnson AE (1986) Proc Natl Acad Sci U S A 83:8604 Kurzchalia TV et al (1986) Nature 320:634 Zopf D, Bernstein HD, Johnson AE, Walter P (1990) EMBO J 9:4511 Connolly T, Rapiejko PJ, Gilmore R (1991) Science 252:1171 Egea PF et al (2004) Nature 427:215 Peluso P et al (2000) Science 288:1640 Powers T, Walter P (1995) Science 269:1422 Cleverley RM, Gierasch LM (2002) J Biol Chem 277:46763 Bourne HR, Sanders DA, Mccormick F (1991) Nature 349:117 Gawronski-Salerno J, Coon JSV, Focia PJ, Freymann DM (2007) Proteins 66:984 Focia PJ, Gawronski-Salerno J, Coon JS, Freymann DM (2006) J Mol Biol 360:631 Freymann DM, Keenan RJ, Stroud RM, Walter P (1999) Nat Struct Biol 6:793 Freymann DM, Keenan RJ, Stroud RM, Walter P (1997) Nature 385:361 Peluso P, Shan SO, Nock S, Herschlag D, Walter P (2001) Biochem 40:15224 Gu SQ et al (2005) RNA-Publ RNA Soc 11:1374 Siu FY, Spanggord RJ, Doudna JA (2007) RNA 13:240 Sagar MB, Lucast L, Doudna JA (2004) RNA 10:772 Brown S, Fournier MJ (1984) J Mol Biol 178:533 Jagath JR et al (2001) RNA-Publ RNA Soc 7:293 Zheng N, Gierasch LM (1997) Mol Cell 1:79 Poritz MA, Strub K, Walter P (1988) Cell 55:4 Poritz MA et al (1990) Science 250:1111 Batey RT, Sagar MB, Doudna JA (2001) J Mol Biol 307:229 Hainzl T, Huang S, Sauer-Eriksson AE (2007) Proc Natl Acad Sci U S A 104:14911 Stroud RM, Walter P (1999) Curr Opin Struct Biol 9:754 Spanggord RJ, Siu F, Ke AL, Doudna JA (2005) Nat Struct Mol Biol 12:1116 Mandon EC, Jiang Y, Gilmore R (2003) J Cell Biol 162:575 Buskiewicz I et al (2005) J Mol Biol 351:417 Powers T, Walter P (1997) EMBO J 16:4880 Connolly T, Gilmore R (1989) Cell 57:599 Rapiejko PJ, Gilmore R (1992) J Cell Biol 117:493 Zopf D, Bernstein HD, Walter P (1993) J Cell Biol 120:1113 Jagath JR, Rodnina MV, Wintermeyer W (2000) J Mol Biol 295:745 Padmanabhan S, Freymann DM (2001) Structure 9:859 Ramirez UD et al (2002) J Mol Biol 320:783 Focia PJ, Alam H, Lu T, Ramirez UD, Freymann DM (2004) Proteins 54:222 Lakowicz JR (1999) Principles of fluorescence spectroscopy.

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Multistate GTPase Control Co-translational Protein Targeting by Xin Zhang
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