This study will investigate the solution structure and folding of bovine myelin basic protein species that have been separated into the nonphosphorylated, one phosphate residue, two phosphate residue and three phosphate residue species. Each one of the four myelin basic protein charge isomers will be reacted with the crosslinking agents 2-(p-nitrophenyl)allyltrimethylammonium iodide and 2-(p-nitrophenyl)allyl-4-nitro-3-carboxyphenyl sulfide. These reagents react with lysine residues via Michael addition to give products of thermodynamic control rather than kinetic control as produced by other crosslinking agents. The intra and inter molecularly crosslinked protein will be separated and the lysine residues involved will be determined via tryptic peptide mapping and amino acid analysis. Parallel experiments using ethylene glycol bis (succinimidyl succinate) to give the products of kinetic control will also be carried out. Analysis of the crosslinking results should permit a determination of the proximal lysine residues in the folded protein. The influence of the phosphate residues on the secondary structure and folding of the protein will be studied using Raman and FT-IR spectroscopy. Also, the effect of pH, ionic strength and solvent on the diffusion coefficient and radius of gyration will be determined using quasielastic light scattering. The results of this study will assist in understanding the role of structure in antigenicity of this protein. In turn that understanding will assist in helping to understand the role of the protein in Multiple Sclerosis and EAE. The data will also assist in understanding the role of this protein in the myelin membrane.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
1R01NS023474-01
Application #
3406991
Study Section
Neurological Sciences Subcommittee 1 (NLS)
Project Start
1986-04-01
Project End
1989-03-31
Budget Start
1986-04-01
Budget End
1987-03-31
Support Year
1
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Type
Organized Research Units
DUNS #
791277940
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109