Limited proteolysis of myelin basic protein in a system mimetic of the myelin interlamellar aqueous space.
We have investigated the early steps of myelin basic protein (MBP) degradation in a membrane mimetic system (reverse micelles), resembling the interlamellar aqueous spaces where the protein is located in the myelin sheath. MBP, unfolded in buffer, refolds on incorporation into the micelles, resulting in reduced accessibility to three proteolytic enzymes, trypsin, cathepsin D, and Staphylococcus aureus V8 protease, in comparison with aqueous solution. Eleven cleavage sites seen in buffer are removed from proteolytic attack in micellar solution. These sites delineate a protected protein domain displaying a potential beta-sheet structure capable of interacting with the myelin membrane. An additional site not seen in buffer is attacked in the micelles. Experiments with a structure inducer, 15% 1-propanol in buffer, reveal that the refolding pattern of MBP in reverse micelles is specific to the membrane biomimetic system and is not produced by organic solvent per se. Micellar digestions of MBP generate long peptides, two of which, isolated after tryptic digestion, have been found to be immunodominant in multiple sclerosis patients. The findings suggest the structure induced in MBP by the micelles resembles that leading to production of the self-peptides recognized by T cells during proteolytic breakdown of MBP in autoimmune demyelinating diseases.
Nicot
C
C
Biomédicale des Saints-Pères, Laboratoire des Systèmes Moléculaires Organisés, Paris, France.
Vacher
M
M
Denoroy
L
L
Kahn
P C
PC
Waks
M
M
D000595:Amino Acid Sequence; D000818:Animals; D002402:Cathepsin D / Q000378:metabolism; D002417:Cattle; D010450:Endopeptidases / Q000378:metabolism*; D008823:Micelles; D008969:Molecular Sequence Data; D004676:Myelin Basic Protein / Q000737:chemistry / Q000378:metabolism*; D009186:Myelin Sheath / Q000378:metabolism*; D011433:Propranolol / Q000494:pharmacology; D017510:Protein Folding; D017433:Protein Structure, Secondary; D012697:Serine Endopeptidases / Q000378:metabolism; D012996:Solutions; D014357:Trypsin / Q000378:metabolism; D014867:Water
D008823:Micelles; D004676:Myelin Basic Protein; D012996:Solutions; D014867:Water; D011433:Propranolol; D010450:Endopeptidases; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D014357:Trypsin; D002402:Cathepsin D