D 2002

The effect of cultivation conditions on protein composition of membrane of Paracoccus denitrificans

BOUCHAL, Pavel and Igor KUČERA

Basic information

Original name

The effect of cultivation conditions on protein composition of membrane of Paracoccus denitrificans

Name in Czech

Vliv kultivačních podmínek na proteinové složení membránové frakce Paracoccus denitrificans

Authors

BOUCHAL, Pavel (203 Czech Republic, guarantor) and Igor KUČERA (203 Czech Republic)

Edition

Brno, VI. Pracovní setkání biochemiků a molekulárních biologů. Sborník příspěvků. p. 67-67, 2002

Publisher

Masarykova univerzita

Other information

Language

English

Type of outcome

Stať ve sborníku

Field of Study

10600 1.6 Biological sciences

Country of publisher

Czech Republic

Confidentiality degree

není předmětem státního či obchodního tajemství

RIV identification code

RIV/00216224:14310/02:00019686

Organization unit

Faculty of Science

ISBN

80-210-2777-0

Keywords in English

Paracoccus denitrificans; membrane proteins; proteome; 2-D electrophoresis
Změněno: 25/5/2006 12:18, doc. Mgr. Pavel Bouchal, Ph.D.

Abstract

V originále

New method using two-dimensional gel electrophoresis was optimised for Paracoccus denitrificans cell envelope study. This approach consist of sample preparation especially optimised for Paracoccus denitrificans membrane proteins, carrier-ampholyte isoelectric focusation in the first dimension (CA-IEF) and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) in the second dimension. Acquired 2-D gels were compared using image analysis and PDQUEST software. Various conditions of initial solubilisation using various detergents were tested. Sodium dodecyl sulphate (SDS), 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfo-nate (CHAPS), dodecyl-â-D-maltoside and 6-aminocaproic acid were used for this purpose. Sample buffer and IEF gel composition effects on 2-D gels resolution were also tested. Extraction using free detergent was compared with extraction using diluted detergent in mixture with sample buffer. Dilution ratio between presolubilised extract and sample buffer was optimised. Focusation algorithm was also optimised for this purpose.

In Czech

Byla optimalizována nová metoda k analýze membránové frakce bakterie Paracoccus denitrificans za použití isoelektrické fokusace s nosnými amfolyty a dodecylsulfátové polyakrylamidové elektroforézy. Při optimalizaci byly testovány různé postupy přípravy vzorku za použití různých detergentů. 2-D gely byly vyhodnocovány pomocí software PDQUEST.

Links

GA203/01/1589, research and development project
Name: Mechanizmy regulace respiračních systémů denitrifikačních bakterií faktory prostředí
Investor: Czech Science Foundation, Mechanisms involved in regulation of respiratory systems in denitrification bacteria by environmental factors
MSM 143100005, plan (intention)
Name: Strukturně-funkční vztahy biomolekul a jejich role v metabolismu
Investor: Ministry of Education, Youth and Sports of the CR, Biomolecular Structure-function Relationships and their role in the Metabolism