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Volumn 38, Issue 4, 1999, Pages 509-512

Chemo-enzymatic synthesis of fluorescent Rab 7 proteins: Tools to study vesicular trafficking in cells

Author keywords

Enzyme catalysis; Fluorescence spectroscopy; Isoprenoids; Lipoproteins; Signal transduction

Indexed keywords

GUANINE NUCLEOTIDE BINDING PROTEIN; RAB PROTEIN;

EID: 0033557870     PISSN: 14337851     EISSN: None     Source Type: Journal    
DOI: 10.1002/(sici)1521-3773(19990215)38:4<509::aid-anie509>3.0.co;2-3     Document Type: Article
Times cited : (34)

References (26)
  • 9
    • 0026800719 scopus 로고
    • Previous studies of the interaction of prenyltransferases with their lipid substrate have been conducted primarily with radiolabeled phosphoisoprenoids or by using the change in the fluorescence of endogenous tryptophane upon substrate binding. See a) M. C. Seabra, M. S. Brown, C. A. Slaughter, T. C. Sudhof, J. L Goldstein, Cell 1992, 70, 1049;
    • (1992) Cell , vol.70 , pp. 1049
    • Seabra, M.C.1    Brown, M.S.2    Slaughter, C.A.3    Sudhof, T.C.4    Goldstein, J.L.5
  • 15
    • 85069762576 scopus 로고
    • b) for reviews on isatoic anhydrides, see G. M. Coppola Synthesis 1980, 505.
    • (1980) Synthesis , pp. 505
    • Coppola, G.M.1
  • 18
    • 33747548563 scopus 로고    scopus 로고
    • note
    • +: 464.1239; found: 464.1294.
  • 19
    • 33747543755 scopus 로고    scopus 로고
    • note
    • +: 532.1865; found: 532.1930.
  • 20
    • 33747565412 scopus 로고    scopus 로고
    • note
    • 13C spectra was observed in the NMR spectra of the diphosphate compounds. This result was found to be concentration and substrate dependent, and is thought to reflect slow rotation about the vinylogous carbamate bonds. The doubling of signals was especially prevalent in the farnesyl derivative 2, and was observed, although to a less degree (ca. 10-15%), in the intermediates that lead up to the farnesyl diphosphate derivative (5b and 6b). The doubling of the NMR signals however, was not observed for the geranyl intermediates (5a and 6a).
  • 22
    • 33747546845 scopus 로고    scopus 로고
    • note
    • 2O and precipitating the protein with 10% trichloroacetic acid. The protein pellet was washed with cold acetone and re-suspended in SDS-PAGE loading buffer. Proteins were resolved on the 15% SDS-PAGE gel and the prenylated proteins were visualized with a Fluor 100 (BioRad) fluorescence scanner.
  • 26
    • 0028040141 scopus 로고
    • RabGGTase baculoviral expression vectors pVL-RabGGTalpha and pVL-RabGGTbeta were obtained from the American Type Culture Collection (order numbers 87154 and 87155, respectively). Baculoviruses were generated and RabGGTase was expressed and purified as described previously: F. P. Cremers, S. A. Armstrong, M. C. Seabra, M. C. Brown, J. L. Goldstein J. Biol. Chem. 1994, 209, 2111.
    • (1994) J. Biol. Chem. , vol.209 , pp. 2111
    • Cremers, F.P.1    Armstrong, S.A.2    Seabra, M.C.3    Brown, M.C.4    Goldstein, J.L.5


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.