Isoelectric focusing of proteins in the native and denatured states. Anomalous behaviour of plasma albumin.
1. An analytical technique of isoelectric focusing in thin layers of polyacrylamide gel has been used to determine the isoelectric point, pI, of several proteins in the presence and in the absence of concentrated urea. 2. The presence of urea did not greatly affect pI except for bovine plasma albumin, where an increase of approx. 1pH unit was found. 3. Evidence is presented that this change in the pI of bovine plasma albumin is due to the normalization of certain ionizable groups on unfolding of the protein in urea. 4. Evidence is also presented that prolonged exposure of bovine plasma albumin to urea results in intramolecular disulphide interchange and that, on removal of urea, the new patterns of disulphide bonding stabilize abnormal conformations with pI values intermediate between those of the native and denatured states. 5. The studies demonstrate heterogeneity in bovine plasma albumin based on primary-sequence differences. 6. Isoelectric focusing of proteins in urea appears to be useful in the study of various aspects of protein structure.
D000478:Alkylation; D001215:Asparaginase; D001536:Bence Jones Protein; D002919:Chymotrypsinogen; D003402:Creatine Kinase; D004586:Electrophoresis; D005033:Ethylmaleimide; D007328:Insulin; D007525:Isoelectric Focusing; D007782:Lactoglobulins; D009211:Myoglobin; D010047:Ovalbumin; D010084:Oxidation-Reduction; D011489:Protein Denaturation; D012710:Serum Albumin, Bovine; D014508:Urea
D007328:Insulin; D007782:Lactoglobulins; D009211:Myoglobin; D012710:Serum Albumin, Bovine; D014508:Urea; D010047:Ovalbumin; D001536:Bence Jones Protein; D002919:Chymotrypsinogen; D003402:Creatine Kinase; D001215:Asparaginase; D005033:Ethylmaleimide