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

被引：19

作者：

Testa, Lorenzo ^{[1
]}

Brocca, Stefania ^{[1
]}

Samalikova, Maria ^{[1
]}

Santambrogio, Carlo ^{[1
]}

Alberghina, Lilia ^{[1
]}

Grandori, Rita ^{[1
]}

机构：

[1] Univ Milano Bicocca, Dept Biosci & Biotechnol, I-20126 Milan, Italy

来源：

BIOTECHNOLOGY JOURNAL | 2011年 / 6卷 / 01期

关键词：

Charge state distributions; Cyclin-dependent protein kinase inhibitors; Nano-electrospray; Partially folded states; Sic1; KINASE INHIBITOR SIC1; ESI MS; DETERMINANTS; ENSEMBLES;

D O I：

10.1002/biot.201000253

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：96 / 100

页数：5

共 20 条

[1] The yeast cyclin-dependent kinase inhibitor Sic1 and mammalian p27Kip1 are functional homologues with a structurally conserved inhibitory domain [J].

Barberis, M ;

De Gioia, L ;

Ruzzene, M ;

Sarno, S ;

Coccetti, P ;

Fantucci, P ;

Vanoni, M ;

Alberghina, L .

BIOCHEMICAL JOURNAL, 2005, 387 :639-647

[2] Co-populated conformational ensembles of β2-microglobulin uncovered quantitatively by electrospray ionization mass spectrometry [J].

Borysik, AJH ;

Radford, SE ;

Ashcroft, AE .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (26) :27069-27077

[3] Defining Structural Domains of an Intrinsically Disordered Protein: Sic1, the Cyclin-Dependent Kinase Inhibitor of Saccharomyces cerevisiae [J].

Brocca, Stefania ;

Testa, Lorenzo ;

Samalikova, Maria ;

Grandori, Rita ;

Lotti, Marina .

MOLECULAR BIOTECHNOLOGY, 2011, 47 (01) :34-42

[4] Order propensity of an intrinsically disordered protein, the cyclin-dependent-kinase inhibitor Sic1 [J].

Brocca, Stefania ;

Samalikova, Maria ;

Uversky, Vladimir N. ;

Lotti, Marina ;

Vanoni, Marco ;

Alberghina, Lilia ;

Grandori, Rita .

PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS, 2009, 76 (03) :731-746

[5] Detection of multiple protein conformational ensembles in solution via deconvolution of charge-state distributions in ESI MS [J].

Dobo, A ;

Kaltashov, IA .

ANALYTICAL CHEMISTRY, 2001, 73 (20) :4763-4773

[6] Biophysical characterization of intrinsically disordered proteins [J].

Eliezer, David .

CURRENT OPINION IN STRUCTURAL BIOLOGY, 2009, 19 (01) :23-30

[7] Natively unfolded proteins [J].

Fink, AL .

CURRENT OPINION IN STRUCTURAL BIOLOGY, 2005, 15 (01) :35-41

[8] Regulation of cell division by intrinsically unstructured proteins: Intrinsic flexibility, modularity, and signaling conduits [J].

Galea, Charles A. ;

Wang, Yuefeng ;

Sivakolundu, Sivashankar G. ;

Kriwacki, Richard W. .

BIOCHEMISTRY, 2008, 47 (29) :7598-7609

[9] Investigation of intact protein complexes by mass spectrometry [J].

Heck, AJR ;

van den Heuvel, RHH .

MASS SPECTROMETRY REVIEWS, 2004, 23 (05) :368-389

[10] The cyclin-dependent kinase inhibitory domain of the yeast Sic1 protein is contained within the C-terminal 70 amino acids [J].

Hodge, A ;

Mendenhall, M .

MOLECULAR AND GENERAL GENETICS, 1999, 262 (01) :55-64

← 1 2 →