RESUMO
Cytochrome and lipid composition of membranes is considered as the attributes required for adaptation of the alkalophiles to alkaline conditions. Respiratory chains of alkalophilic representatives of the genus Bacillus are discussed. Special attention is paid to the features of the Na(+)-cycle of these bacteria and to the features determining halo- and alkalotolerant phenotype, which have been reported due to recent achievements in genomics.
Assuntos
Bacillus/genética , Bacillus/metabolismo , Metabolismo Energético , Citocromos/metabolismo , Transporte de Elétrons , Genômica , Metabolismo dos Lipídeos , Modelos Biológicos , Oxirredução , Fenótipo , Sódio/química , Sódio/metabolismoRESUMO
Site-mutations were introduced into putative cation binding site 1 of the H,K-ATPase at glu-797, thr-825, and glu-938. The side chain oxygen of each was not essential but the mutations produced different activation and inhibition kinetics. Site mutations thr-825 (ala, leu) and glu-938 (ala, gln) modestly decreased the apparent affinity to K+, while glu-797 (gln) was equivalent to wild type. As expected of competitive inhibition, mutations of thr-825 and glu-938 that decreased the apparent affinity for K+ also increased the apparent affinity for SCH28080. This is consistent with the participation of thr-825 and glu-938 in a cation binding domain. The sidechain geometry, but not the sidechain charge of glu-797, is essential to ATPase function as the site mutant glu-797 (gly) inactivated the H,K-ATPase, while glu-797 (gln) was active but the apparent affinity to SCH 28080 was decreased by four-fold. Lys-793, a unique residue of the H,K-ATPase, was essential for ATPase function. Since this residue is adjacent to site 1, the result suggests that charge pairing between lys-793 and residues at or near this site may be essential to ATPase function.
Assuntos
ATPase Trocadora de Hidrogênio-Potássio/metabolismo , Estômago/enzimologia , Animais , Sítios de Ligação/genética , ATPases Transportadoras de Cálcio/química , ATPases Transportadoras de Cálcio/metabolismo , Cátions/metabolismo , ATPase Trocadora de Hidrogênio-Potássio/química , ATPase Trocadora de Hidrogênio-Potássio/genética , Proteínas de Membrana/genética , Modelos Químicos , Mutagênese Sítio-Dirigida , Potássio/metabolismo , Proto-Oncogenes , Coelhos , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismoRESUMO
The culture, morphology, and genome of alkalo- and halotolerant bacterial strain Bacillus sp. FTU were characterized; the strain is compared to other representatives of genus Bacillus. The DNA-DNA hybridization data indicate that the strains of Bacillus halodurans DSM 497 and DSM 2513 and Bacillus sp. FTU belong to the same species. Bacillus sp. FTU can be renamed to Bacillus halodurans FTU. The N-terminal amino acid fragments of the subunits I and II of the terminal caa3-type cytochrome c oxidase of B. halodurans FTU were sequenced. The N-terminal fragments of this enzyme and of the caa3-type oxidase of alkalophilic Bacillus firmus OF4 are highly homologous (homology of subunits I and II is over 90 and over 96%, respectively). Such high homology of the terminal oxidases of these bacteria might be due to their alkaline medium.
Assuntos
Bacillus/enzimologia , Bacillus/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Sequência de Aminoácidos , Bacillus/classificação , DNA Bacteriano/genética , Complexo IV da Cadeia de Transporte de Elétrons/química , Dados de Sequência Molecular , Conformação Proteica , Homologia de Sequência de Aminoácidos , Especificidade da EspécieRESUMO
The interaction of caldesmon with certain Ca-binding proteins was investigated by means of electrophoresis under non-denaturating conditions. In the presence of Ca2+ calmodulin, troponin C and S-100 protein form a complex with caldesmon. No complex formation takes place in the absence of Ca2+. Lactalbumin and pike parvalbumin (pI4.2) do not interact with caldesmon independently of Ca-concentration. Both S-100 protein and calmodulin effectively inhibit phosphorylation of caldesmon by Ca-phospholipid-dependent protein kinase. At low ionic strength S-100 protein reverses the inhibitory action of caldesmon on the skeletal muscle acto-heavy meromyosin ATPase more effectively than calmodulin. It is supposed that in certain tissues and cell compartments the proteins belonging to the S-100 family are able to substitute for calmodulin in the caldesmon-dependent regulation of actin and myosin interaction.
