title: Deimination of myelin basic protein. 1. Effect of deimination of arginyl residues of myelin basic protein on its structure and susceptibility to digestion by cathepsin D.
abstract: The effect of deimination of arginyl residues in bovine myelin basic protein (MBP) on its susceptibility to digestion by cathepsin D has been studied. Using bovine component 1 (C-1) of MBP, the most unmodified of the components with all 18 arginyl residues intact, we have generated a number of citrullinated forms by treatment of the protein with purified peptidylarginine deiminase (PAD) in vitro. We obtained species containing 0-9.9 mol of citrulline/mol of MBP. These various species were digested with cathepsin D, a metalloproteinase which cleaves proteins at Phe-Phe linkages. The rate of digestion compared to component 1 was only slightly affected when 2.7 or 3.8 mol of citrulline/mol of MBP was present. With 7.0 mol of citrulline/mol of MBP, a large increase in the rate of digestion occurred. No further increase was observed with 9.9 mol of citrulline/mol of MBP. The immunodominant peptide 43-88 (bovine sequence) was released slowly when 2.7 and 3.8 mol of citrulline/mol of MBP was present, but it was released rapidly when 7.0 mol of citrulline/mol of MBP was present. The dramatic change in digestion with 7.0 mol of citrulline/mol of MBP or more could be explained by a change in three-dimensional structure. Molecular dynamics simulation showed that increasing the number of citrullinyl residues above 7 mol/mol of MBP generated a more open structure, consistent with experimental observations in the literature. We conclude that PAD, which deiminates arginyl residues in proteins, decreases both the charge and compact structure of MBP. This structural change allows better access of the Phe-Phe linkages to cathepsin D. This scheme represents an effective way of generating the immunodominant peptide which sensitizes T-cells for the autoimmune response in demyelinating disease.
authors.affiliation: Department of Structural Biology & Biochemistry, The Hospital for Sick Children, Toronto, Ontario, Canada M5G 1X8.
authors.forename: L B
authors.initials: LB
authors.lastname: Pritzker
authors.forename: S
authors.initials: S
authors.lastname: Joshi
authors.forename: J J
authors.initials: JJ
authors.lastname: Gowan
authors.forename: G
authors.initials: G
authors.lastname: Harauz
authors.forename: M A
authors.initials: MA
authors.lastname: Moscarello
mesh_terms: D000818:Animals; D001120:Arginine / Q000737:chemistry*; D002403:Cathepsins / Q000378:metabolism*; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D002956:Citrulline / Q000737:chemistry; D020278:Demyelinating Autoimmune Diseases, CNS / Q000201:enzymology / Q000209:etiology; D006867:Hydrolases / Q000378:metabolism; D007700:Kinetics; D013058:Mass Spectrometry; D008958:Models, Molecular; D004676:Myelin Basic Protein / Q000737:chemistry*; D010446:Peptide Fragments / Q000276:immunology / Q000378:metabolism; D011487:Protein Conformation; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases
chemical_list: D004676:Myelin Basic Protein; D010446:Peptide Fragments; D002956:Citrulline; D001120:Arginine; D006867:Hydrolases; D002403:Cathepsins; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases