Escherichia coli ilvN interacts with the FAD binding domain of ilvB and activates the AHAS I enzyme

Biochemistry
Ashima Mitra, S Sarma

Abstract

The unique multidomain organization in the multimeric Escherichia coli AHAS I (ilvBN) enzyme has been exploited to generate polypeptide fragments which, when cloned and expressed, reassemble in the presence of cofactors to yield a catalytically competent enzyme. Multidimensional multinuclear NMR methods have been employed for obtaining near complete sequence specific NMR assignments for backbone HN, 15N, 13Calpha and 13Cbeta atoms of the FAD binding domain of ilvB on samples that were isotopically enriched in 2H, 13C and 15N. Unambiguous assignments were obtained for 169 of 177 backbone Calpha atoms and 127 of 164 side chain Cbeta atoms. The secondary structure determined on the basis of observed 13Calpha secondary chemical shifts and sequential NOEs agrees well with the structure of this domain in the catalytic subunit of yeast AHAS. Binding of ilvN to the ilvBalpha and ilvBbeta domains was studied by both circular dichroism and isotope edited solution nuclear magnetic resonance methods. Changes in CD spectra indicate that ilvN interacts with ilvBalpha and ilvBbeta domains of the catalytic subunit and not with the ilvBgamma domain. NMR chemical shift mapping methods show that ilvN binds close to the FAD binding site in ilvBbet...Continue Reading

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Citations

Apr 13, 2010·Biomolecular NMR Assignments·N Megha Karanth, Siddhartha P Sarma
Mar 21, 2013·Chembiochem : a European Journal of Chemical Biology·Yuefang ZhaoZhen Xi
Sep 1, 2016·Applied Microbiology and Biotechnology·Yadi LiuXiaoyuan Wang
Aug 8, 2015·Scientific Reports·Chunxu SongJos M Raaijmakers
Aug 7, 2017·MicrobiologyOpen·Yadi LiuXiaoyuan Wang
Sep 19, 2018·Chembiochem : a European Journal of Chemical Biology·Yonghui XieZhen Xi
Oct 19, 2013·Nature Reviews. Neuroscience·Gaute T EinevollStefano Panzeri
Jun 17, 2020·The Biochemical Journal·Yingying ZhangYuequan Shen

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