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Visual detection of specific, native interactions between soluble and microbead-tethered alpha-helices from membrane proteins


, : Visual detection of specific, native interactions between soluble and microbead-tethered alpha-helices from membrane proteins. Biochemistry 40(46): 13753-9

Using peptides tethered to polymer microbeads, we have developed a technique for measuring the interactions between the transmembrane alpha-helices of membrane proteins and for screening combinatorial libraries of peptides for members that interact with specific helices from membrane proteins. The method was developed using the well-characterized homodimerization sequence of the membrane-spanning alpha-helix from the erythrocyte membrane protein glycophorin A (GPA). As a control, we also tested a variant with a dimer-disrupting alteration of a critical glycine residue to leucine. To test for detectable, native interactions between detergent-solubilized and microbead-tethered alpha-helices, we incubated fluorescent dye-labeled GPA analogues in sodium dodecyl sulfate solution with microbeads that contained covalently attached GPA analogues. When the dye-labeled peptide in solution and the bead-tethered peptide both contained the native glycophorin A sequence, the microbeads readily accumulated the dye through lateral peptide-peptide interactions and were visibly fluorescent under UV light. When either the peptide in solution or the peptide attached to the beads contained the glycine to leucine change, the beads did not accumulate any dye. The usefulness of this method for screening tethered peptide libraries was tested by incubating dye-labeled, native sequence peptides in detergent solution with a few native sequence beads plus an excess of beads containing the variant glycine to leucine sequence. When the dye-labeled peptide in solution was present at a concentration of > or =2 microM, the few native sequence beads were visually distinguishable from the others because of their bright fluorescence. Using this model system, we have shown that it is possible to visually detect specific, native interactions between alpha-helices from membrane proteins using peptides tethered to polymer microbeads. It will thus be possible to use this method to measure the specific lateral interactions that drive the folding and organization of membrane proteins and to screen combinatorial libraries of peptides for members that interact with them.

US$29.90

PMID: 11705363


Other references

Hammond, A., 1972: Dysplasia epiphysialis, with ocular anomalies. Ophthalmologica. Journal International D'ophtalmologie. International Journal of Ophthalmology. Zeitschrift für Augenheilkunde 165(6): 554-555

Bahk; Chang; Cheon; Cho; Jang; Hahn; Hara; Lim; Kang; Kim; Kim; Kim; Kim; Kim; Kim; Koo; Lee; Lim; Moon; Nam; Pac; Park; Park; Ryu; Shin; Sim; Song; Woo; Yokoyama; Yoon, 1991: Diffractive excitation of 14.6-, 60-, and 200-GeV/nucleon 16O and 14.6-GeV/nucleon 28Si nuclei in nuclear emulsion. Physical Review. C, Nuclear Physics 43(3): 1410-1419

Ng, T.P.; Mak, K.H.; Phua, K.H.; Tan, C.H., 1999: Trends in mortality, incidence, hospitalisation, cardiac procedures and outcomes of care for coronary heart disease in Singapore, 1991-1996. In this study, we used Singapore population-based data from 1991 to 1996 to examine recent trends in mortality, incidence and hospitalisation for acute myocardial infarction (AMI), and explored the roles of primary prevention and medical care inte...

Uzura, A.; Katsuragi, T.; Tani, Y., 2001: Optimal conditions for production of (R)-1-phenylpropanol by Fusarium moniliforme strain MS31. Resting cells of Fusarium moniliforme strain MS31 produced (R)-1-phenylpropanol from propylbenzene. The components of the medium and the reaction conditions were adjusted to increase the specific activity of the hydroxylating enzyme involved. Gluc...

Francetti, L.; Maggiore, E.; Marchesi, A.; Ronchi, G.; Romeo, E., 1991: Oral hygiene in subjects treated with diphenylhydantoin: effects of a professional program. The purpose of the present study was to investigate, on a longitudinal basis, the effectiveness of a specific preventive dental program for patients who are taking phenytoin for seizure control. The results confirm that a preventive dental program...

Cecil, R.L.; Vaughan, H.F., 1919: Results Of Prophylactic Vaccination Against Pneumonia At Camp Wheeler. 1. 13,460 men, or about 80 per cent of the entire camp strength, were vaccinated against pneumonia with pneumococcus lipovaccine. 2. The dosage employed in all cases was 1 cc. of the lipovaccine containing approximately 10 billion each of Pneumoco...

Goldwasser, R.A.; Kemron, A.; Nobel, T.A., 1961: Fluorescent antibody in the diagnosis of rabies. Brain specimens were received from 424 animals of 22 species suspected of rabies. The fluorescent antibody technique, using fluores-cein isothiocyanate, gave 41 positive results, 40 of which were confirmed by inoculation of mice or staining for Ne...

Berry, J.D.; Henny, C.J., 1995: Osprey polygyny in Wyoming. Journal of Raptor Research 29(4): 279-281

Pound, A.W., 1966: Further observations concerning the influence of preliminary stimulation by croton oil and acetic acid on the initiation of skin tumours in mice by urethane. British Journal of Cancer 20(2): 385-398

Marroux, H.J.B.; Orr-Ewing, A.J., 2016: Distinguishing Population and Coherence Transfer Pathways in a Metal Dicarbonyl Complex Using Pulse-Shaped Two-Dimensional Infrared Spectroscopy. Collection of two-dimensional infrared (2DIR) spectra using two ultrafast, broadband infrared pump pulses followed by an ultrafast probe pulse optimizes the experimental time and frequency resolution, but can also introduce quantum beat and cohere...