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Aulabaugh A Ding W Kapoor B Tabei K Alksne L Dushin R Zatz T Ellestad G Huang X 《Analytical biochemistry》2007,360(1):14-22
Many bacterial surface proteins containing an LPXTG motif are anchored to the cell wall peptidoglycan by catalysis with the thiol transpeptidase sortase. The transpeptidation and hydrolysis reactions of sortase have been proposed to proceed through a common acyl enzyme intermediate. The reactions of Staphylococcus aureus sortase with fluorogenic substrate Abz-LPETG-Dnp in the presence or absence of triglycine were characterized in this study to gain additional insight into the kinetic mechanism of sortase. We report here the development of a reverse-phase HPLC assay to identify and characterize sortase reaction intermediates. The HPLC results provide for the first time clear evidence for the formation of a kinetically competent acyl enzyme intermediate during the overall transpeptidation reaction. The results also suggest that sortase undergoes an unexpected intramolecular acyl transfer reaction in the absence of a nucleophile. The significance of this type of HPLC assay as a tool to study enzyme mechanism is discussed. 相似文献
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Sabrina Degaspari Carmen Branco Tzanno-Martins Clarice Kazue Fujihara Roberto Zatz Jo?o Paulo Branco-Martins Tania Araujo Viel Hudson de Souza Buck Ana Maria Marques Orellana Ana Elisa B?hmer Larissa de Sá Lima Diana Zukas Andreotti Carolina Demarchi Munhoz Cristoforo Scavone Elisa Mitiko Kawamoto 《PloS one》2015,10(5)
Renal insufficiency can have a negative impact on cognitive function. Neuroinflammation and changes in klotho levels associate with chronic kidney disease (CKD) and may play a role in the development of cognitive impairment (CI). The present study evaluates the correlation of cognitive deficits with neuroinflammation and soluble KLOTHO in the cerebral spinal fluid (CSF) and brain tissue of nephrectomized rats (Nx), with 5/6 renal mass ablation. Nx and sham Munich Wistar rats were tested over 4 months for locomotor activity, as well as inhibitory avoidance or novel object recognition, which started 30 days after the surgery. EMSA for Nuclear factor-κB and MILLIPLEXMAP or ELISA kit were used to evaluate cytokines, glucocorticoid and KLOTHO levels. Nx animals that showed a loss in aversive-related memory and attention were included in the CI group (Nx-CI) (n=14) and compared to animals with intact learning (Nx-M n=12 and Sham n=20 groups). CSF and tissue samples were collected 24 hours after the last behavioral test. The results show that the Nx-groups have increased NF-κB binding activity and tumor necrosis factor-alpha (TNF-α) levels in the hippocampus and frontal cortex, with these changes more pronounced in the Nx-CI group frontal cortex. In addition, the Nx-CI group showed significantly increased CSF glucocorticoid levels and TNF-α /IL-10 ratio compared to the Sham group. Klotho levels were decreased in Nx-CI frontal cortex but not in hippocampus, when compared to Nx-M and Sham groups. Overall, these results suggest that neuroinflammation mediated by frontal cortex NF-κB, TNF-α and KLOTHO signaling may contribute to Nx-induced CI in rats. 相似文献
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M Zatz R B Levisky J A Levy B O Valente M Gianotti O Frota-Pessoa 《Journal de génétique humaine》1973,21(4):297-305
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M Zatz 《Life sciences》1977,21(9):1267-1276
Cholera toxin, which activates adenylate cyclase by a mechanism independent of the β-adrenergic receptor, induced more N-acetyltransferase activity in supersensitive than in subsensitive rat pineal glands. This was due to an increased response in supersensitive glands, of several of the components involved in the induction. Although there was no difference in the number of binding sites for cholera toxin, there was more toxin-stimulated adenylate cyclase activity in the supersensitive glands. There was also a larger accumulation of cyclic AMP and a greater stimulation of protein kinase activity in the supersensitive glands. It is inferred that changes in the number of β-adrenergic binding sites are not the primary basis for changes in sensitivity. Rather, there are multiple sites of regulation, possibly affected by a common mechanism. 相似文献
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Electrical field stimulation caused the release of preloaded 3H-norepinephrine and of cyclic GMP from the rat pineal gland. Increased release of catecholamines and of cyclic nucleotide occurred at low frequency and current, and was largely dependent on the presence of intact nerve endings and extracellular calcium. Presynaptic synthesis and release of cyclic GMP appears to accompany the exocytotic release of neurotransmitter. 相似文献
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Jean Pierre Schatzmann Peron Auriléia Aparecida de Brito Mayra Pelatti Wesley Nogueira Brand?o Luana Beatriz Vitoretti Flávia Regina Greiffo Elaine Cristina da Silveira Manuel Carneiro Oliveira-Junior Mariangela Maluf Lucila Evangelista Silvio Halpern Marcelo Gil Nisenbaum Paulo Perin Carlos Eduardo Czeresnia Niels Olsen Saraiva Camara Flávio Aimbire Rodolfo de Paula Vieira Mayana Zatz Ana Paula Ligeiro de Oliveira 《PloS one》2015,10(9)