2004
The effects of terminal electron acceptor and sodium azide on the Paracoccus denitrificans protein composition
BOUCHAL, Pavel, Petra PŘECECHTĚLOVÁ, Zbyněk ZDRÁHAL a Igor KUČERAZákladní údaje
Originální název
The effects of terminal electron acceptor and sodium azide on the Paracoccus denitrificans protein composition
Název česky
Vliv terminálního akceptoru elektronů a azidu na proteinové složení bakterie Paracoccus denitrificans
Autoři
BOUCHAL, Pavel (203 Česká republika, garant), Petra PŘECECHTĚLOVÁ (203 Česká republika), Zbyněk ZDRÁHAL (203 Česká republika) a Igor KUČERA (203 Česká republika)
Vydání
Olomouc, Acta Universitatis Palackianae Olomucensis - Chemica 43S, od s. 28-29, 2 s. 2004
Nakladatel
Palackého univerzita Olomouc
Další údaje
Jazyk
angličtina
Typ výsledku
Stať ve sborníku
Obor
10600 1.6 Biological sciences
Stát vydavatele
Česká republika
Utajení
není předmětem státního či obchodního tajemství
Kód RIV
RIV/00216224:14310/04:00010926
Organizační jednotka
Přírodovědecká fakulta
ISBN
80-244-0882-1
Klíčová slova anglicky
Paracoccus denitrificans; proteome analysis; terminal electron acceptor; azide
Změněno: 18. 2. 2005 14:24, doc. Mgr. Pavel Bouchal, Ph.D.
V originále
Paracoccus denitrificans is a non-fermentative, facultatively autotrophic soil bacterium often studied in the field of bioenergetics, particularly due to resemblance of its aerobic respiratory chain to that of mitochondria. Also an aspect of a great nutritional adaptability was discovered, related to the ability of exploiting various electron donors and electron acceptors for maintenance and growth. To discuss mechanisms underlying regulation of gene expression by growth conditions, a high-throughput proteome mapping is one of the most effective tools to-date. For 2-D electrophoretic analysis of whole-cell extract and its membrane fraction, isoelectric focusation with immobilized pH gradients in the first dimension and SDS-PAGE in the second dimension were used. Using this approach, more than 800 protein spots of total cell extract and membrane fraction were detected in the range pI 3-10 and Mr 15-100 kDa. We compared complex protein composition of whole cells of P. denitrificans cultivated under aerobic and various anaerobic conditions (with nitrate, nitrite and nitrous oxide). We also investigated the effect of respiratory inhibitor azide on proteomic profiles of the cells grown aerobically and anaerobically with NO2-. The similar approach was applied also on membrane fractions of cells (grown on O2, O2+N3-, NO3-). The most distinct proteomic profile matches the growth on nitrous oxide; the probable reasons are discussed. The significant changes in protein composition of total cell lysates were caused also by azide. Popis výsledku (dříve anotace) v anglickém jazyce (rozsah pro RIV: max. 1000 znaků)ÚdajeAbout 50 protein spots have been submitted to the analysis by peptide mass fingerprinting using MALDI-TOF MS. However, the genome of P. denitrificans has not been completely sequenced yet and, currently, information about only 98+126 proteins is available from Swiss?Prot/TrEMBL database. Due to this fact, we were able to identify only 8 proteins up to-date. Among them, nitrite reductase and succinate dehydrogenase are key enzymes in P. denitrificans metabolism. Nitrite reductase is the second of four enzymes catalyzing the denitrification pathway in P. denitrificans, where NO3- is reduced via NO2-, NO, N2O to molecular nitrogen under anaerobic conditions. Succinate dehydrogenase, fumarate-producing enzyme in citrate cycle, was previously found to be very close to the bovine heart mitochondrial enzyme, having four subunits (64.9, 28.9, 13.4 and 12.5 kDa). Our ID corresponds with the 28.9 kDa FeS subunit. We also compared expression data of nitrite reductase obtained by proteome analysis with the measurement of nitrite reductase enzyme activity. These data were in a good agreement. The quantitative data and protein maps are kept in a database in PDQUEST format. The web-accessible proteome 2-D database has been established at http://www.mpiib-berlin.mpg.de/2D-PAGE.
Česky
Proteomová analýza bakterie Paracoccus denitrificans v závislosti na použitém terminálním akceptoru elektronů a přítomnosti azidu. Jedná se o předběžnou prezentaci proteomové studie, která byla následně publikována: Bouchal P., Přecechtělová P., Zdráhal Z., Kučera I.: Protein composition of Paracoccus denitrificans cells grown on various electron acceptors and in the presence of azide. Proteomics 2004, 4, 2662-2671.
Návaznosti
| GA203/01/1589, projekt VaV |
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| MSM 143100005, záměr |
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| MSM 143100008, záměr |
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