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Volume 75, Issue 4, 30 May 2008, Pages 1041-1045   

title:
A coupled reagent of o-phthalaldehyde and sulfanilic acid for protein detection based on the measurements of light scattering signals with a common spectrofluorometer
author:Yuan Fang Li, Xiao Wei Shena and Cheng Zhi Huang
College of Chemistry and Chemical Engineering, CQKL-RATA, Southwest University, Chongqing 400715, China


Received 17 September 2007;  revised 1 January 2008;  accepted 2 January 2008.  Available online 16 January 2008

Abstract
The covalent binding of pyridoxal 5¡ä-phosphate (PLP) to human serum albumin (HSA) is important in the regulation of PLP metabolism. In plasma, PLP is bound to HSA at a single high-affinity and at two or more nonspecific sites. To characterize the primary PLP binding site, HSA was incubated with [3H]PLP, and the Schiff base linkage was reduced with potassium borohydride. Tryptic peptides were purified, and the major labeled peptide was sequenced. Amino acid analysis confirmed a homogeneous peptide Leu-Asp-Glu-Leu-Arg-Asp-Glu-Gly-Xaa-Ala-Ser-Ser-Ala-Lys which corresponds to residues 182¨C195 of HSA. The data in...
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