Analysis of self-association of bovine neurophysins by gel chromatography.
Analytical gel chromatography has been used to examine self-association of bovine neurophysins I and II under several sets of conditions. The data provide no evidence for associated species larger than the dimer. Association constants and Stokes radii of both monomer and dimer are very similar for both proteins in both 0.1 M KOAc, 0.16 M KCl and 0.1 M KPO4, 0.16 M KCl at pH 5.6 and 25 degrees C. The average values derived for the Stokes radii of the monomer and dimer under these conditions are 14.5 +/- 0.7 and 23.0 +/- 0.4 A, respectively. These results confirm the conclusion of Rholam and Nicolas [(1981) Biochemistry 20, 5837-5843] that the monomer and, to a lesser extent, the dimer are highly assymmetric. The Stokes radius of the monomer calculated by Rholam and Nicolas (op cit.) is approximately 30% larger than the value derived here. This discrepancy is probably the result of end-on penetration of the gel by elongated molecules [Y. Nozaki, N. M. Schechter, J. A. Reynolds, and C. Tanford (1976) Biochemistry 15, 3884-3890]. In contrast to Tellam and Winzor [(1980) Arch. Biochem. Biophys. 201, 20-24], it was found that neurophysin II does not exist solely as the dimer in 0.1 M KPO4, pH 5.6, although substitution of 0.1 M KPO4 for 0.1 M KOAc does increase the association constant by a factor of seven. Addition of 1.4 M LiCl at pH 8.1 also increases the association constant sevenfold, as well as increasing the Stokes radius of the monomer approximately 20%. The effects of ionic strength are consistent with the conclusion of Nicolas et al. [(1978) J. Biol. Chem 253, 2633-2639] that formation of the dimer depends upon hydrophobic bonding.
D000818:Animals; D002021:Buffers; D002417:Cattle; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002712:Chlorides / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008094:Lithium / Q000494:pharmacology; D018021:Lithium Chloride; D046911:Macromolecular Substances; D008970:Molecular Weight; D009481:Neurophysins; D009994:Osmolar Concentration; D010710:Phosphates / Q000494:pharmacology; D011188:Potassium / Q000494:pharmacology; D017680:Potassium Compounds
D002021:Buffers; D002712:Chlorides; D046911:Macromolecular Substances; D009481:Neurophysins; D010710:Phosphates; D017680:Potassium Compounds; D008094:Lithium; C013216:potassium phosphate; D018021:Lithium Chloride; D011188:Potassium