Cyst(e)ine residues of bovine white-matter proteolipid proteins. Role of disulphides in proteolipid conformation.
Cyst(e)ine residues of bovine white-matter proteolipid proteins were characterized in a highly purified preparation. From a total of 10.6 cyst(e)ine residues/molecule of protein, as determined by performic acid oxidation, 2.5-3 thiol groups were freely accessible to iodoacetamide, iodoacetic acid and 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB), when the proteins were solubilized in chloroform/methanol (C/M) (2:1, v/v). The presence of lipids had no effect on thiol-group exposure. One thiol group available to DTNB in C/M could not be detected when proteolipids were solubilized in the more polar solvent n-butanol. In a C/M solution of purified proteolipid proteins, SDS did not increase the number of reactive thiol groups, but the cleavage of one disulphide bridge made it possible to alkylate six more groups. C.d. and fluorescence studies showed that rupture of this disulphide bond changed the protein conformation, which was reflected in partial loss of helical structure and in a greater exposure to the solvent of at least one tryptophan residue. Cyst(e)ine residues were also characterized in the different components [PLP (principal proteolipid protein), DM20 and LMW (low-Mr proteins)] of the proteolipid preparation. Although the numbers of cyst(e)ine residues in PLP and DM20 were similar, in LMW fewer residues were alkylated under four different experimental conditions. The differences, however, are not simply related to differences in Mr.
Oteiza
P I
PI
Departamento de Quimica Biologica, Facultad de Farmacia y Bioquimica, Universidad de Buenos Aires, Argentina.
Adamo
A M
AM
Aloise
P A
PA
Paladini
A C
AC
Paladini
A A
AA
Soto
E F
EF
D000818:Animals; D002417:Cattle; D002942:Circular Dichroism; D003545:Cysteine / Q000032:analysis*; D003553:Cystine / Q000032:analysis*; D004220:Disulfides / Q000032:analysis; D004228:Dithionitrobenzoic Acid; D005561:Formates; D007461:Iodoacetates; D009417:Nerve Tissue / Q000032:analysis*; D011487:Protein Conformation; D011510:Proteolipids; D013050:Spectrometry, Fluorescence
D004220:Disulfides; D005561:Formates; D007461:Iodoacetates; D011510:Proteolipids; C008817:peroxyformic acid; D003553:Cystine; D004228:Dithionitrobenzoic Acid; D003545:Cysteine