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Volumn 286, Issue 5440, 1999, Pages 782-785

Microtubule disassembly by ATP-dependent oligomerization of the AAA enzyme katanin

Author keywords

[No Author keywords available]

Indexed keywords

ADENOSINE TRIPHOSPHATASE; ADENOSINE TRIPHOSPHATE; ALPHA TUBULIN; BETA TUBULIN; MONOMER; NUCLEOTIDE; PROTEIN KINASE P60;

EID: 0033595814     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.286.5440.782     Document Type: Article
Times cited : (197)

References (39)
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    • note
    • em 508) using either an SLM 8100 in photon counting mode (Spectronics) or a Perkin-Elmer LS-5B (Perkin-Elmer). Standards and NSF D2 were quantitated by the Bradford assay.
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    • note
    • ECFP (Clonetech) and EYFP (Clonetech) coding sequences were inserted into the pFastBac-p60 expression plasmid as in (15).
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    • note
    • Amino acid residues are abbreviated as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; Y, Tyr.
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    • note
    • The E334Q mutant was prepared by overlap extension amplification [S. N. Ho et al., Gene 77, 51 (1989)] of the wild-type p60 cDNA, followed by cloning of the mutant sequence into pFastBac HT B as in (15) and sequencing to check for amplification errors.
  • 26
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    • note
    • 2, 0.02% Triton X-100, 5% glycerol, and 1 mM ATP. Standard proteins were BSA (Stokes radius 35 Å), catalase (52 Å), and thyroglobulin (86 Å). We estimated molecular mass by using the measured Stokes radius and the S value [L M. Siegel and K. J. Monty, Biochim. Biophys. Acta 112, 346 (1966)].
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    • em 508).
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    • 2-terminus of GFP.
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    • note
    • We performed ATPase assays by using the malachite green method as described in (7). We included DEAE-purified ATP at 1 mM.
  • 32
    • 0344130085 scopus 로고    scopus 로고
    • note
    • We also examined FRET when the katanin concentration was varied with a constant microtubule concentration in the presence of ATP-γ-S. The signal did not increase in a hyperbolic manner. Instead, an increase in the FRET signal was observed only when the katanin reached a critical concentration, which again suggests a cooperative phenomenon.
  • 33
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    • note
    • c2+ depolymerized)]/microtubule concentration, μM).
  • 39
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    • note
    • We thank F. J. McNally, R. D. Mullins, K. S. Thorn, and J. E. Wilhelm for critical reading of the manuscript.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.