التفاصيل البيبلوغرافية
| العنوان: |
A Novel Substrate-Binding Site in the X-ray Structure of an Oxidized E. coli Glyceraldehyde 3-Phosphate Dehydrogenase Elucidated by Single-Wavelength Anomalous Dispersion. |
| المؤلفون: |
Annia, Rodríguez-Hernández, Romo-Arévalo, Enrique, Rodríguez-Romero, Adela |
| المصدر: |
Crystals (2073-4352); Dec2019, Vol. 9 Issue 12, p622-622, 1p |
| مصطلحات موضوعية: |
GLYCERALDEHYDEPHOSPHATE dehydrogenase, RECOMBINANT proteins, DISPERSION (Chemistry), CATALYTIC domains, X-rays |
| مستخلص: |
Escherichia coli (E. coli), one of the most frequently used host for the expression of recombinant proteins, is often affected by the toxic effect of the exogenous proteins that is required to express. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is a multi-functional protein that has been used as a control marker for basal function and it is known to undergo cysteine oxidation under different types of cellular stress. Here, we report the 3D structure of the endogenous GAPDH purified from stressed E. coli cells expressing a eukaryotic protein. The structure was solved at 1.64 Å using single-wavelength anomalous dispersion (SAD) phasing with a selenium-modified enzyme. Interestingly, each GAPDH monomer contains a molecule of glyceraldehyde-3 phosphate in a non-previously identified site. Furthermore, the catalytic Cys149 is covalently attached to a ~300 Da molecule, possibly glutathione. This modification alters the conformation of an adjacent alpha helix in the catalytic domain, right opposite to the NAD+ binding site. The conformation of the alpha helix is stabilized after soaking the crystals with NAD+. These results exemplify the effects that the overexpression of an exogenous protein has over the host proteins and sheds light on the structural changes that large oxidant molecules on the catalytic cysteine produce for the GAPDH enzyme. [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
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