4.5 Article

Electrospray ionization-mass spectrometry conformational analysis of isolated domains of an intrinsically disordered protein

期刊

BIOTECHNOLOGY JOURNAL
卷 6, 期 1, 页码 96-100

出版社

WILEY-V C H VERLAG GMBH
DOI: 10.1002/biot.201000253

关键词

Charge state distributions; Cyclin-dependent protein kinase inhibitors; Nano-electrospray; Partially folded states; Sic1

资金

  1. MIUR-Italbionet
  2. University Milano-Bicocca (Fondo Ateneo per la Ricerca)

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The highly dynamic and heterogeneous molecular ensembles characterizing intrinsically disordered proteins (IDP) in solution pose major challenges to the conventional methods for structural analysis. Electrospray ionization-mass spectrometry (ESI-MS) allows direct detection of distinct conformational components, effectively capturing also partially folded and short-lived states. We report the description of two complementary fragments (1-186 and 187-284) of the IDP Sic1, a cyclin-dependent protein kinase inhibitor of yeast Saccharomyces cerevisiae. Structural heterogeneity is noted in both cases, but the two fragments reveal slightly different conformational properties. The results are consistent with previously reported differences between the two protein moieties and corroborate the feasibility of IDP conformational analysis by ESI-MS.

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