Metal-dependent hydrolysis of myelin basic protein by IgGs from the sera of patients with multiple sclerosis.
Homogeneous IgG fractions were obtained by chromatography of the sera of patients with multiple sclerosis (MS) on Protein G-Sepharose under conditions that remove non-specifically bound proteins. These IgGs contained several chelated metals, the relative amount of which decreases in the order: Fe>or=Ca>Cu>or=Zn>or=Mg>or=Mn>or=Pb>or=Co>or=Ni. In contrast to homogeneous IgGs of healthy individuals, Abs of MS patients effectively hydrolyzed human myelin basic protein (MBP). A minor metal-dependent fraction was obtained by chromatography of highly purified IgGs from MS patient on Chelex-100. This IgG fraction did not hydrolyze human MBP in the absence of Me(2+) ions but was activated after addition of Me(2+) ions: Mg(2+)>Mn(2+)>Cu(2+)>Ca(2+). Proteolytic activities of IgGs from other MS patients were also activated by other metal ions (Ni(2+), Fe(2+), Co(2+), Zn(2+), Pb(2+), and Co(2+)) and especially Ni(2+). Ni(2+)-activated IgGs were separated into distinct MBP-hydrolyzing fractions by chromatography on HiTraptrade mark Chelating Sepharose charged with Ni(2+). Detection of Mg(2+)-dependent proteolytic activity in the SDS-PAGE area corresponding only to IgG provided direct evidence that IgG from sera of MS patients possesses metal-dependent human MBP-hydrolyzing activity. Observed properties of MS abzymes distinguish them from other known mammalian metalloproteases and demonstrate their pronounced catalytic diversity. Metal-dependent IgGs from MS patients represent the first example of abzymes with metal-dependent proteolytic activity.
Polosukhina
Dar'ya I
DI
Institute of Chemical Biology and Fundamental Medicine, Siberian Division of Russian Academy of Sciences, Novosibirsk, Russia.
Kanyshkova
Tat'yana G
TG
Doronin
Boris M
BM
Tyshkevich
Olga B
OB
Buneva
Valentina N
VN
Boiko
Alexey N
AN
Gusev
Evgenii I
EI
Nevinsky
Georgy A
GA
Favorova
Olga O
OO
D000293:Adolescent; D000328:Adult; D017151:Antibodies, Catalytic / Q000378:metabolism* / Q000494:pharmacology; D005260:Female; D006801:Humans; D006868:Hydrolysis; D008297:Male; D008673:Metals, Alkaline Earth / Q000494:pharmacology*; D019216:Metals, Heavy / Q000494:pharmacology*; D008875:Middle Aged; D009103:Multiple Sclerosis / Q000097:blood / Q000276:immunology*; D004676:Myelin Basic Protein / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*
D017151:Antibodies, Catalytic; D008673:Metals, Alkaline Earth; D019216:Metals, Heavy; D004676:Myelin Basic Protein