Reversible binding interactions between the tryptophan enantiomers and albumins of different animal species as determined by novel high performance liquid chromatographic methods: an attempt to localize the D- and L-tryptophan binding sites on the human serum albumin polypeptide chain by using protein fragments.
The stereoselectivity of the reversible binding interactions between the D- and L-tryptophan enantiomers and serum albumins of different animal species and fragments of human serum albumin (HSA) was investigated by applying three novel high performance liquid chromatographic (HPLC) arrangements. The separations were performed by means of 1) an achiral (diol-bond), 2) a chiral (bovine serum albumin-bond) silica gel sorbent, and 3) a column switching technique which uses both the diol- and HSA-bond HPLC stationary phases. A polarimetric detector and/or an ultraviolet (UV) spectrophotometer were used to monitor the separation process. HPLC arrangement 3 allowed the evaluation of enantioselective binding for D- and L-tryptophan to different albumins and albumin fragments. At present, column switching can be considered the technique of the broadest applicability for investigating the reversible binding interactions between a protein and drug enantiomers.
Soltés
L
L
Institute of Experimental Pharmacology, Slovak Academy of Sciences, Bratislava, Slovak Republic.
Sebille
B
B
D000818:Animals; D001665:Binding Sites; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D004285:Dogs; D006168:Guinea Pigs; D006736:Horses; D006801:Humans; D007202:Indicators and Reagents; D010446:Peptide Fragments / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D011048:Polarography; D011817:Rabbits; D017459:Receptors, Amino Acid / Q000737:chemistry*; D012709:Serum Albumin / Q000737:chemistry* / Q000378:metabolism; D012756:Sheep; D013045:Species Specificity; D013056:Spectrophotometry, Ultraviolet; D013237:Stereoisomerism; D013552:Swine; D014364:Tryptophan / Q000737:chemistry*
D007202:Indicators and Reagents; D010446:Peptide Fragments; D010455:Peptides; D017459:Receptors, Amino Acid; D012709:Serum Albumin; D014364:Tryptophan