The physiochemical and immunological characteristics of rat lymph binding protein for 25-hydroxyvitamin D3.
The binding proteins for 25-hydroxyvitamin D3(25-OH-D3) in rat lymph and plasma were purified to apparent homogeneity as determined by sodium dodecyl sulfate disc gel electrophoresis. The purification procedures included the following techniques: gel filtration on Sephadex G-100 and G-200; affinity chromatography on Blue Sepharose CL-6B; ion-exchange chromatography on DEAE-Sepharose CL-6B; chromatofocussing on a Mono P column. Both proteins from lymph and plasma were eluted with the same retention time from an isoelectric column at a pH of approximately 4.53 and showed nearly identical data on the analysis of amino acid composition. When specific anti-lymph 25-OH-D3-binding protein antiserum was prepared in a rabbit, and Ouchterlony immunodiffusion was performed, the same precipitate line was observed on both lymph and plasma binding proteins. These results strongly suggest that a common transport protein exists in both rat lymph and plasma and circulates in the blood-lymph system.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002112:Calcifediol / Q000378:metabolism*; D002352:Carrier Proteins / Q000097:blood / Q000276:immunology / Q000302:isolation & purification*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002851:Chromatography, High Pressure Liquid; D000939:Epitopes / Q000276:immunology; D005779:Immunodiffusion; D008196:Lymph / Q000032:analysis*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D014809:Vitamin D-Binding Protein
D000596:Amino Acids; D002352:Carrier Proteins; D000939:Epitopes; D014809:Vitamin D-Binding Protein; D002112:Calcifediol