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Specific Binding of GAPDH in the Presence or Absence of Selenium to SPS Monitored by Affinity Chromatography
Author(s) -
GallowayYunusah Michelle,
Stadtman Thressa C.
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.1027.2
Subject(s) - glyceraldehyde 3 phosphate dehydrogenase , selenium , biochemistry , cysteine , chemistry , selenoprotein , enzyme , affinity chromatography , selenocysteine , dehydrogenase , glutathione , glutathione peroxidase , organic chemistry
The methods of synthesis and insertion of selenium into selenocysteine (Sec)_containing enzymes has been recognized in prokaryotes, eukaryotes and archaebacteria. Yet, the transport and intracellular trafficking of selenium is incompletely characterized. Therefore, considerable attention has reallocated to the identification and characterization of selenium adducts or delivery proteins required in specific metabolic pathways. It has been documented the Escherichia coli glyceraldehyde_3_phosphate dehydrogenase (GAPDH; EC ) actively bound selenium possibly as a perselenide derivative following in vivo labeling with 75Se[selenite] [Lacourciere, G.M., Levine, R.L. and Stadtman, T.C. (2002) PNAS USA 99 , 9150_9153]. Subsequently, Yuki and coworkers [Ogasawara, Y., Lacourciere, G.M., Ishii, K. and Stadtman, T.C. (2005) PNAS USA 102 , 1012_1016] provided in vitro evidence that each subunit of human erythrocyte GAPDH possessed a reactive cysteine residue, which can act as the binding site for selenium supplied as selenodiglutathione from selenite and glutathione. These findings suggest GAPDH participated as a component of a selenium delivery system for the biosynthesis of selenophosphate, a reactive selenium compound, catalyzed by the E. coli selenophosphate synthetase (SPS). To date, there are no reports relating to the binding capability of SPS to native and 75 Se_labeled GAPDH. Consequently, the work presented herein reports the application of affinity chromatography as a screening tool to emphasize the specificity of the Se enzymes. This research was fully supported by the National Heart, Lung, and Blood Institute, National Institutes of Health.