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Volumn 306, Issue 5698, 2004, Pages 1042-1046

Tracking SNARE complex formation in live endocrine cells

Author keywords

[No Author keywords available]

Indexed keywords

CATALYST ACTIVITY; CELL MEMBRANES; COMPLEXATION; CYTOLOGY; FLUORESCENCE; NEUROLOGY;

EID: 8344255773     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.1102559     Document Type: Article
Times cited : (117)

References (33)
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  • 13
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    • note
    • 2 (17). The parallel orientation of ΔSN2 in such complexes would bring CFP into proximity not with Venus but with another CFP, possibly favoring nonfluorescent energy transfer between two CFPs. The complex would also hinder the approach of CFP and Venus that might occasionally occur by thermal flexing of unstructured SCORE. Both effects would tend to diminish FRET.
  • 15
    • 8344275065 scopus 로고    scopus 로고
    • note
    • All constructs were found not only in the plasma membrane but also inside the cell, especially when strongly overexpressed. However, the proportion of SCORE inside cells did not vary systematically between different SCORE mutants (fig. S1).
  • 20
    • 8344257943 scopus 로고    scopus 로고
    • note
    • In the very dimmest cells, autofluorescence resulted in an apparent FRET ratio of 1 with all three probes. At higher probe concentrations, the influence of autofluorescence waned, as documented by the Syx-insensitive probe SCORE ΔSN1, whose FRET values approached those seen under other conditions (Fig. 3, C and F).
  • 24
    • 0037033997 scopus 로고    scopus 로고
    • K. Hu et al., Nature 415, 646 (2002).
    • (2002) Nature , vol.415 , pp. 646
    • Hu, K.1
  • 29
    • 8344250897 scopus 로고    scopus 로고
    • note
    • Purified SCORE (10 μM) was pulled up into a glass microcuvette (20-μm path length, Vitro Dynamics, New York), whose ends were then sealed with vaseline. The cuvette was placed under a drop of water and imaged as if it were a cell.
  • 31
    • 8344233347 scopus 로고    scopus 로고
    • note
    • 2+].
  • 32
    • 2142803437 scopus 로고    scopus 로고
    • Materials and methods are available as supporting material on Science Online.
    • Science Online
  • 33
    • 8344263009 scopus 로고    scopus 로고
    • note
    • We thank J. Adelman, C. Bond, and S. Bae for help. Supported by NIH MH60600. Molecular interaction data have been deposited in the Biomolecular Interaction Network Database with accession codes 180392 and 180393.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.