Electrospray ionization mass spectrometry analysis of the apo‐ and metal‐substituted forms of the Fur protein

FEBS Letters

Jaquinod, Michel³  Latour, Jean-Marc¹  Michaud-Soret, Isabelle¹  Forest, Eric³  Touati, Danièle²  Adrait, Annie¹ 

[1] Département de Recherche Fondamentale sur la Matière Condensée, Service de Chimie Inorganique et Biologique, Laboratoire de Chimie de Coordination (Unité de Recherche Associée au CNRS No. 1194), CEA-Grenoble, 38054 Grenoble, France;Institut Jacques Monod, CNRS, Université Paris 7, 75251 Paris Cedex 05, France;Laboratoire de Spectrométrie de Masse des Protéines, Institut de Biologie Structurale, 38027 Grenoble Cedex 1, France

关键词: Ferric uptake regulation (Fur) protein;    Iron uptake regulation;    Electrospray ionization mass spectrometry;    Escherichia coli;    Fur protein;    Ferric Uptake Regulation protein;    Co-Fur and Mn-Fur;    Fur protein substituted with cobalt and manganese ions;    respectively;    Fur-Met;    Fur protein without the N-terminal methionine;    ESI-MS;    electrospray ionization-mass spectrometry;    MALDI-TOF;    matrix-assisted-laser desorption/ionization-time of flight;    HPLC;    high pressure liquid chromatography;    LC;    liquid chromatography;    ICP-AES;    inductive coupling plasma-atomic emission spectroscopy;    PMSF;    phenylmethylsulfonyl fluoride;   

DOI  :  10.1016/S0014-5793(97)00963-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Fur has been purified and reconstituted with Co²⁺ and Mn²⁺. The ESI-MS spectra of the apoprotein as well as Mn-Fur and Co-Fur under acidic denaturating conditions showed the existence of two species of molecular mass 16 660±3 and 16 792±3 Da, which correspond, respectively, to the N-terminal methionine `excised' or `non-excised' forms of the monomer. This result proves the absence of any other post-translational modification or modification due to metal incorporation. On the other hand, under soft conditions, ESI spectra provided for the first time direct evidence for dimeric metal-containing forms in solution.

【 授权许可】

Unknown