Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1979 Nov;76(11):5460–5464. doi: 10.1073/pnas.76.11.5460

Photoreactive labeling of M13 coat protein in model membranes by use of a glycolipid probe.

V W Hu, B J Wisnieski
PMCID: PMC411668  PMID: 293655

Abstract

Coliphage M13 coat protein in synthetic bilayer membranes was labeled by use of 12-(4-azide-2-nitrophenoxy)stearoyl[1-14C]glucosamine, a photoreactive glycolipid probe that spontaneously inserts into membranes. In this model system, the probe preferentially labeled the proteins over the lipids. Experiments designed to test the probe's restriction to integral membrane proteins revealed that extrinsic proteins as well as external peptide fragments of integral membrane proteins were not accessible to the photogenerated nitrene on the fatty acid chain. Only integral membrane peptides were labeled by the membrane-bound probe. These results indicate that protein labeling can be effected specifically from within the hydrocarbon milieu of a model membrane system.

Full text

PDF
5463

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  2. Bramhall J. S., Shiflett M. A., Wisnieski B. J. Mapping the membrane proteins of Newcastle-disease virus with a photoreactive glycolipid probe. Biochem J. 1979 Feb 1;177(2):765–768. doi: 10.1042/bj1770765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bramhall J., Ishida B., Wisnieski B. Photolabile and paramagnetic reagents for the investigation of transmembrane signaling events. J Supramol Struct. 1978;9(3):399–406. doi: 10.1002/jss.400090310. [DOI] [PubMed] [Google Scholar]
  4. Chakrabarti P., Khorana G. A new approach to the study of phospholipid-protein interactions in biological membranes. Synthesis of fatty acids and phospholipids containing photosensitive groups. Biochemistry. 1975 Nov 18;14(23):5021–5033. doi: 10.1021/bi00694a001. [DOI] [PubMed] [Google Scholar]
  5. Iwata K. K., Manweiler C. A., Bramhall J., Wisnieski B. J. Photoreactive probes for high resolution mapping of membrane proteins. Prog Clin Biol Res. 1978;22:579–589. [PubMed] [Google Scholar]
  6. Knippers R., Hoffmann-Berling H. A coat protein from bacteriophage fd. I. Hydrodynamic measurements and biological characterization. J Mol Biol. 1966 Nov 14;21(2):281–292. doi: 10.1016/0022-2836(66)90099-4. [DOI] [PubMed] [Google Scholar]
  7. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  8. Stoffel W., Salm K., Körkemeier U. Biosynthetic incorporation of fatty acids with photosensitive groups into membrane lipids of cells in tissue culture. Hoppe Seylers Z Physiol Chem. 1976 Jul;357(7):917–924. doi: 10.1515/bchm2.1976.357.2.917. [DOI] [PubMed] [Google Scholar]
  9. Wickner W. Asymmetric orientation of a phage coat protein in cytoplasmic membrane of Escherichia coli. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4749–4753. doi: 10.1073/pnas.72.12.4749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Wickner W. Asymmetric orientation of phage M13 coat protein in Escherichia coli cytoplasmic membranes and in synthetic lipid vesicles. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1159–1163. doi: 10.1073/pnas.73.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Wisnieski B. J., Bramhall J. S. Labeling of the active subunit of cholera toxin from within the membrane bilayer. Biochem Biophys Res Commun. 1979 Mar 15;87(1):308–313. doi: 10.1016/0006-291x(79)91680-2. [DOI] [PubMed] [Google Scholar]
  12. Wisnieski B. J., Iwata K. K. Electron spin resonance evidence for vertical asymmetry in animal cell membranes. Biochemistry. 1977 Apr 5;16(7):1321–1326. doi: 10.1021/bi00626a013. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES