Determining membrane protein topology using fluorescence protease protection (FPP)

Journal of Visualized Experiments

Volumn 2015, Issue 98, 2015, Pages

  White, Carl ^a   Nixon, Alex ^a   Bradbury, Neil A ^a  

^a ROSALIND FRANKLIN UNIVERSITY OF MEDICINE AND SCIENCE   (United States)

Author keywords

Cellular biology; Digitonin; Fluorescence assay; GFP; Issue 98; Membrane protein; Protease; Proteolysis; Topology

Indexed keywords

DIGITONIN; GREEN FLUORESCENT PROTEIN; HYBRID PROTEIN; MEMBRANE PROTEIN; PEPTIDE HYDROLASE;

CELL MEMBRANE; CHEMISTRY; CYTOSOL; ENDOPLASMIC RETICULUM; FLUORESCENCE MICROSCOPY; HEK293 CELL LINE; HUMAN; METABOLISM; MITOCHONDRION; PROCEDURES;

CELL MEMBRANE; CYTOSOL; DIGITONIN; ENDOPLASMIC RETICULUM; GREEN FLUORESCENT PROTEINS; HEK293 CELLS; HUMANS; MEMBRANE PROTEINS; MICROSCOPY, FLUORESCENCE; MITOCHONDRIA; PEPTIDE HYDROLASES; RECOMBINANT FUSION PROTEINS;

EID: 84941243012     PISSN: 1940087X     EISSN: None     Source Type: Journal    
DOI: 10.3791/52509     Document Type: Article

References (25)

1. Sevier, C. S., & Kaiser, C. A. Formation and transfer of disulphide bonds in living cells. Nat Rev Mol Cell Biol. 3, 836-847, doi: 10.1038/nrm954 (2002).
2. Brodsky, J. L., & Skach, W. R. Protein folding and quality control in the endoplasmic reticulum: Recent lessons from yeast and mammalian cell systems. Current Opinion In Cell Biology. 23, 464-475, doi: 10.1016/j.ceb.2011.05.004 (2011).
3. Kyte, J., & Doolittle, R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 157, 105-132 (1982).
4. Ott, C. M., & Lingappa, V. R. Integral membrane protein biosynthesis: why topology is hard to predict. J Cell Sci. 115, 2003-2009 (2002).
5. Traxler, B., Boyd, D., & Beckwith, J. The topological analysis of integral cytoplasmic membrane proteins. The Journal of Membrane Biology. 132, 1-11 (1993).
6. Cserzo, M., Wallin, E., Simon, I., von Heijne, G., & Elofsson, A. Prediction of transmembrane alpha-helices in prokaryotic membrane proteins: the dense alignment surface method. Protein Eng. 10, 673-676 (1997).
7. Geest, M., & Lolkema, J. S. Membrane topology and insertion of membrane proteins: search for topogenic signals. Microbiol Mol Biol Rev. 64, 13-33 (2000).
8. Bogdanov, M., Zhang, W., Xie, J., & Dowhan, W. Transmembrane protein topology mapping by the substituted cysteine accessibility method (SCAM(TM)): application to lipid-specific membrane protein topogenesis. Methods. 36, 148-171, doi:10.1016/j.ymeth.2004.11.002 (2005).
9. Chang, X. B., Hou, Y. X., Jensen, T. J., & Riordan, J. R. Mapping of cystic fibrosis transmembrane conductance regulator membrane topology by glycosylation site insertion. J Biol Chem. 269, 18572-18575 (1994).
10. Hu, C. D., Chinenov, Y., & Kerppola, T. K. Visualization of interactions among bZIP and Rel family proteins in living cells using bimolecular fluorescence complementation. Mol Cell. 9, 789-798 (2002).
11. Zamyatnin, A. A., et al. Assessment of the integral membrane protein topology in living cells. Plant J. 46, 145-154, doi: 10.1111/ j.1365-313X.2006.02674.x (2006).
12. Lorenz, H., Hailey, D. W., & Lippincott-Schwartz, J. Fluorescence protease protection of GFP chimeras to reveal protein topology and subcellular localization. Nat Methods. 3, 205-210, doi: 10.1038/nmeth857 (2006).
13. Sambrook, J., Fritsch, E. F., & Maniatis, T. Molecular Cloning. a Laboratory Manual. 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).
14. Shah, K., McCormack, C. E., & Bradbury, N. A. Do you know the sex of your cells? American journal of physiology. Cell Physiology. 306, C3-C18, doi: 10.1152/ajpcell.00281.2013 (2014).
15. Wang, H., & Brautigan, D. L. A novel transmembrane Ser/Thr kinase complexes with protein phosphatase-1 and inhibitor-2. J Biol Chem. 277, 49605-49612, doi: 10.1074/jbc.M209335200 (2002).
16. Nixon, A., Jia, Y., White, C., & Bradbury, N. A. Fluorescence protease protection reveals the topology of the integral membrane protein Lemur Tyrosine Kinase 2 (LMTK2). Am J Physiol. 304 (2), C164-169, doi: 10.1152/ajpcell.00288.2012 (2012).
17. Tagawa, A., et al. Assembly and trafficking of caveolar domains in the cell: caveolae as stable, cargo-triggered, vesicular transporters. J Cell Biol. 170, 769-779, doi: 10.1083/jcb.200506103 (2005).
18. Rizzuto, R., et al. A gene specifying subunit VIII of human cytochrome c oxidase is localized to chromosome 11 and is expressed in both muscle and non-muscle tissues. J Biol Chem. 264, 10595-10600 (1989).
19. Rizzuto, R., Brini, M., Pizzo, P., Murgia, M., & Pozzan, T. Chimeric green fluorescent protein as a tool for visualizing subcellular organelles in living cells. Curr Biol. 5, 635-642 (1995).
20. Lorenz, H., Windl, O., & Kretzschmar, H. A. Cellular phenotyping of secretory and nuclear prion proteins associated with inherited prion diseases. J Biol Chem. 277, 8508-8516, doi: 10.1074/jbc.M110197200 (2002).
21. Ben-Gedalya, T., et al. Cyclosporin-A-induced prion protein aggresomes are dynamic quality-control cellular compartments. J Cell Sci. 124, 1891-1902, doi: 10.1242/jcs.077693 (2011).
22. Rogers, M., et al. Epitope mapping of the Syrian hamster prion protein utilizing chimeric and mutant genes in a vaccinia virus expression system. J Immunol. 147, 3568-3574 (1991).
23. Lisenbee, C. S., Karnik, S. K., & Trelease, R. N. Overexpression and mislocalization of a tail-anchored GFP redefines the identity of peroxisomal ER. Traffic. 4, 491-501, doi: 10.1034/j.1600-0854.2003.00107.x (2003).
24. Brandee, A. P., et al. Mislocalization and degradation of human P23H-Rhodopsin-GFP in knockin mouse model of retinitis pigmentosa. Invest Ophthalmol Vis Sci. 52, 9728-9736, doi: 10.1167/iovs.11-8654 (2011).
25. Kuznetsov, A. V., et al. Analysis of mitochondrial function in situ in permeabilized muscle fibers, tissues and cells. Nat Protoc. 3, 965-976, doi: 10.1038/nprot.2008.61 (2008).