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Volumn 5, Issue 10, 2005, Pages 1910-1914

Size sorting of protein assemblies using polymeric gradient surfaces

Author keywords

[No Author keywords available]

Indexed keywords

POLYETHYLENE GLYCOLS; POLYMERS; SILICON;

EID: 27544434039     PISSN: 15306984     EISSN: None     Source Type: Journal    
DOI: 10.1021/nl051235h     Document Type: Article
Times cited : (62)

References (42)
  • 22
    • 0030773824 scopus 로고    scopus 로고
    • Howard, J. Nature 1997, 389, 561-567.
    • (1997) Nature , vol.389 , pp. 561-567
    • Howard, J.1
  • 38
    • 27544478028 scopus 로고    scopus 로고
    • note
    • To quantify the amount of surface-bound GFP-kinesin via fluorescence measurements, we used silicon wafers with a 75-nm oxide layer (obtained from GeSiM mbH, Rossendorf) in order to avoid destructive interference effects present at low oxide thickness.
  • 39
    • 27544435066 scopus 로고    scopus 로고
    • note
    • Before usage, the coverslips were immersed in a PEG solution that consisted of 5 mM 2-[methoxypoly-(ethyleneoxy)propyl]-trimethoxysilane (90%, ABCR Karlsruhe) and 0.08% hydrochloric acid (36%) in toluene. After 12 h, the coverslips were rinsed and sonicated in toluene, ethanol, and water. The PEG layer on the glass coverslip prevented kinesin and microtubule binding to that side of the flow cell.
  • 41
    • 27544462688 scopus 로고    scopus 로고
    • note
    • 2, 1 mM Mg-GTP, and 5% DMSO at 37 °C. After 30 min, the microtubule polymers were stabilized and 100-fold diluted in room-temperature BRB80 containing 10 μM taxol. Microtubules were sheared by passing them up and down three times through a 30-gauge needle.
  • 42
    • 27544501166 scopus 로고    scopus 로고
    • note
    • Although, using the presented method the fraction of the short microtubules would always contain a number of long ones, this could potentially be overcome by flowing the microtubules into the device at high concentration. Because of a higher diffusion coefficient, shorter microtubules would reach the active kinsein binding sites faster than the longer ones. Having bound to the surface at high density, the short microtubules would then prevent the binding of the longer ones because of steric hindrance (note that microtubules are stiff polymers with a persistence length of about 1 mm in solution and that they have to land rather parallel to the surface in order to attach to the kinsein motors).


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.