Search results

Search:
in the keywords: Marine actinomycetes

Sort By:

Limit search:

SCREENING THE SPECIFIC SUBSTRATES OF ADENYLATION DOMAIN FROM MARINE ACTINOMYCETES BY FLUORESCENCE QUENCHING AND ISOTHERMAL TITRATION CALORIMETRY

100%

LEI M., ZHAO Z., LIU K., LIU Y., ZHANG H., WU X., GONG S., MA Y., ZHAO H., LIU J., MIN J., QI C.

Acta Poloniae Pharmaceutica - Drug Research

2018

vol. 75

issue 6

1287-1292

Adenylation domain (A domain) is a model of non-ribosomal peptide synthetases (NRPs), responsible for binding and activating the substrates-amino acids. In this study, a pGEX-2T-sare0718 recombinant plasmid (the gene sare0718 was cloned from Salinispora arenicola CNS-205, S.arenicola CNS-205) was transformed and expressed as a protein GST-Sare0718. Fluorescence quenching (FQ) was conducted to investigate the binding of 20 common amino acids to GST-Sare0718, then isothermal titration calorimetry (ITC) also be used[changed]. The results of FQ revealed that intrinsic fluorescence of GST-Sare0718 is quenched steadily by addition of aspartic acid (Asp) and glutamine (Gln) through static quenching mechanism. The binding constants Ka are Asp (1.504×106 L/mol) ≥ Gln (1.468×105 L/mol). And there is one binding site on the protein GST-Sare0718. We confirmed Asp preference of GST-Sare0718 by ITC. Therefore, Asp is the specific substrate. In addition, the experimental data indicates that the prediction system, “the specificity-conferring code”, is not suitable for marine actinomycetes. So it is urgent to set up a special predictive system for marine actinomycetes.

Refine search results

1 Acta Poloniae Pharmaceutica - Drug Research

1 2018

1 GONG S.

1 LEI M.

1 LIU J.

1 LIU K.

1 LIU Y.

1 MA Y.

1 MIN J.

1 QI C.

1 WU X.

1 ZHANG H.

1 ZHAO H.

1 ZHAO Z.

Search results

SCREENING THE SPECIFIC SUBSTRATES OF ADENYLATION DOMAIN FROM MARINE ACTINOMYCETES BY FLUORESCENCE QUENCHING AND ISOTHERMAL TITRATION CALORIMETRY