title: Pseudo-affinity chromatographic approach to probe heterogeneity in buffalo pituitary luteinizing hormone: probable pseudolectin-like behavior of immobilized Cibacron Blue 3GA.
abstract: The alpha (α) and beta (β) subunits of buffalo pituitary luteinizing hormone (LH) were chromatographed on Cibacron Blue 3GA agarose and their immunoreactivity was quantitated using anti-α and anti-β anti sera. Subsequent analyses showed α subunits were relatively more hydrophilic than β subunits. Further, the naturally occurring free α and β subunits were more hydrophobic than their native counterparts which were dissociated and isolated from heterodimeric LH. The lesser sugar content in freely occurring α and beta subunits may be attributed for increased hydrophobicity and consequent upon the existence of their uncombined free forms. In order to ascertain putative sugar-dye interaction, crude LH carrying free subunits, pure LH, and non-glycosylated recombinant β subunit of LH were loaded separately on Cibacron Blue. Methyl mannoside was able to elute 33% of the bound protein in case of crude and pure LH, whereas there was little (3%) elution in case of recombinant LH β subunit. This study suggests a compositional heterogeneity in free and native subunits of LH from the buffalo pituitary. In addition, our findings reveal the pseudolectin-like behavior of Cibacron Blue.
authors.affiliation: Department of Zoology, University of Delhi, Delhi, India.
authors.forename: Taruna
authors.initials: T
authors.lastname: Arora
authors.forename: Nidhi
authors.initials: N
authors.lastname: Vashistha
authors.forename: Rajesh
authors.initials: R
authors.lastname: Chaudhary
authors.forename: Kambadur
authors.initials: K
authors.lastname: Muralidhar
mesh_terms: D000645:Ammonium Sulfate / Q000737:chemistry; D000818:Animals; D000906:Antibodies / Q000378:metabolism; D002020:Buffaloes / Q000378:metabolism*; D002846:Chromatography, Affinity / Q000379:methods*; D007118:Immunoassay / Q000379:methods*; D007986:Luteinizing Hormone / Q000737:chemistry / Q000378:metabolism*; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D014227:Triazines / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000906:Antibodies; D021122:Protein Subunits; D011994:Recombinant Proteins; D014227:Triazines; C012842:Cibacron Blue F 3GA; D007986:Luteinizing Hormone; D000645:Ammonium Sulfate