Identification and characterization of heat shock protein 27 protein species in human myocardial two‐dimensional electrophoresis patterns

Electrophoresis - Tập 18 Số 15 - Trang 2823-2831 - 1997
Christian Scheler1, Eva‐Christina Müller2, Joachim Stahl2, Ursula Müller‐Werdan3, Johann Salnikow1, Peter R. Jungblut4
1Technische Universität Berlin, Institut für Biochemie, Berlin, Germany
2Max Delbrück Centrum für Molekulare Medizin, Berlin, Germany
3Martin‐Luther‐Universität Halle‐Wittenberg, Lehrstuhl für Kardiologische Intensivmedizin an der Klinik für Innere Medizin, Halle, Germany
4Core Facilities / Proteinanalysis, Max Planck Institute for Infection Biology, Max Planck Society

Tóm tắt

AbstractImmunostaining of heat shock protein 27 (Hsp27) protein species on two‐dimensional electrophoresis (2‐DE) gels with enhanced sensitivity yields 59 spots reacting with anti‐Hsp27 antibodies. Recombinant Hsp27 exists in 2‐DE as two major protein species which comigrate in the human myocardial pattern with Hsp27 spots C754 and D899 as defined in the heart high‐performance 2‐DE database (http://www.mdc‐berlin.de/∼emu/heart/). Preparative electrophoresis of human myocardial proteins and analysis of the enriched mass range 20–30 kDa by 2‐DE revealed eight protein spots (C438, C582, C658, C697, C754, C595, C750) from the human myocardial database and a new spot not previously detected on silver‐stained gels. These spots were identified as Hsp27 protein species by enzymatic in‐gel‐digestion and analysis by matrix assisted laser desorption‐ionization (MALDI) peptide mass fingerprinting and, in part, MALDI‐post source decay sequencing of single fragments. Possible post‐translational modifications were investigated: immunostaining tests with anti‐phospho‐serine/‐threonine/‐tyrosine antibodies, although positive for other myocardial proteins, were negative for presumed Hsp27 protein species; likewise, periodate‐glycostaining assays and biotinylation screening did not detect modifications in the investigated Hsp27 protein species.

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Tài liệu tham khảo

10.1002/elps.1150110702

Klose J., 1975, Humangenetik, 26, 211, 10.1007/BF00281458

10.1016/S0021-9258(19)41496-8

10.1002/elps.11501601175

10.1002/elps.1150091208

10.1007/BF01024960

10.1016/S0021-9258(17)35652-1

10.1111/j.1432-1033.1985.tb09157.x

10.1016/S0021-9258(18)61070-1

10.1111/j.1432-1033.1988.tb14308.x

10.1002/elps.1150150159

10.1002/elps.1150160151

10.1021/ac9507956

10.1002/elps.1150170330

Jungblut P. Thiede B. Rev. Mass Spectrom.1997 in press.

10.1002/elps.1150171107

10.1002/elps.1150150197

10.1038/227680a0

10.1002/elps.1150110709

10.1002/elps.1150180524

10.1002/elps.1150171027

10.1016/S0021-9258(19)36654-2

Otto A., 1994, J. Prot. Chem., 13, 478

10.1002/elps.11501301198

10.1002/elps.11501401178

10.1002/elps.11501601355

10.1093/nar/14.10.4127

10.1093/nar/18.21.6457

10.1016/S0021-9258(18)98746-6

10.1016/S0021-9258(17)37091-6

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Electrophoresis

Tập 18 Số 15

2823-2831

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