title: Isoelectric focusing studies of concanavalin A and the lentil lectin.
abstract: Isoelectric focusing (IEF) of metallized and demetallized preparations of concanavalin A (Con A) consisting of either intact or fragmented subunits shows different band patterns. Metallized Con A consisting of intact polypeptide chains (intact Con A) has an isoelectric point (pI) 8.35. Metallized preparations consisting of fragmented chains (fragmented Con A) show three bands with pI values 8.0, 7.8 and 7.7. Demetallized intact Con A (intact apoCon A) has a pI of 6.5, however, it undergoes pH dependent association during IEF under certain conditions, which gives rise to multiple bands. Ampholyte-mediated demetallization of intact and fragmented Con A and subsequent aggregation of the apoprotein results in multiple bands during IEF in the presence of the pH range 3 to 10 ampholytes. However, ampholytes of the pH range 7 to 9 do not demetallize the proteins and show a single band with intact Con A. The pI of intact Con A remains essentially the same in the presence of inhibitory sugar. Furthermore, different moleculars forms of Con A, including locked and unlocked conformers of intact apoCon A, and the dimeric and tetramic states of both intact Con A and intact apoCon A have been identified and their pI values determined. IEF of the lentil isoelectins, LcH-A and LcH-B, shows single bands of pI 8.5 and 9.0, respectively. However, the native lectin mixture gives rise to an additional band of pI 8.8 due to a hybrid protein formed by ampholyte-mediated subunit exchange between the isolectins.
authors.affiliation: Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, NY 10461.
authors.forename: L
authors.initials: L
authors.lastname: Bhattacharyya
authors.forename: C F
authors.initials: CF
authors.lastname: Brewer
mesh_terms: D000665:Ampholyte Mixtures; D003208:Concanavalin A / Q000032:analysis*; D007525:Isoelectric Focusing / Q000379:methods*; D037102:Lectins / Q000032:analysis*; D037121:Plant Lectins; D011487:Protein Conformation
chemical_list: D000665:Ampholyte Mixtures; D037102:Lectins; D037121:Plant Lectins; C033619:lentil lectin; D003208:Concanavalin A