An electrospray ionisation mass spectrometry (ESI‐MS) study to probe the metal ion binding site in the DNA binding domain of the yeast transcriptional activator GAL4

FEBS Letters

Gadhavi, Paresh L¹ 

[1] Cambridge Centre for Molecular Recognition, Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QW, UK

关键词: GAL4;    DNA binding;    Electrospray ionisation mass spectrometry;    Metal binding;   

DOI  :  10.1016/S0014-5793(97)01272-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Electrospray ionisation mass spectrometry (ESI-MS) is used to detect metal ions and their stoichiometry of binding in the DNA binding domain of GAL4. In this analysis, the mass spectra of the apo- and metallo-proteins differ by both mass and charge, precluding the possibility of random adduct formation. Deuterium exchange NMR experiments of Zn(II)-GAL4(7–49) indicate that the binuclear metal ion structure, which is shown to have a net negative charge of −2, is the recipient of several hydrogen bonds, notably from the main-chain amide protons of the ligating cysteine residues, indicating the charge is stabilised in this manner.

【 授权许可】

Unknown