Catalytic properties of IgMs with amylolytic activity isolated from patients with multiple sclerosis.
BACKGROUND
Recently, amylolytic activity was detected in IgMs isolated from the sera of the patients with multiple sclerosis.


METHODS
All purified samples of IgM were electrophoretically homogenous and did not contain any co-purified a-amylase and a-glucosidase activities, in accordance with a set of criteria developed for abzymes. The amylolytic activity of abzymes was studied in the hydrolysis of p-nitrophenyl a-D-maltooligosaccharides with different degrees of polymerization from 1 to 8 by TLC and reverse-phase HPLC techniques.


RESULTS
All IgM samples isolated from 54 patients with clinically definite multiple sclerosis demonstrated hydrolytic activity towards the above artificial substrates. The Michaelis constant values (Km) in the hydrolysis of p-nitrophenyl a-D-maltoheptaoside were in the range of 10 p-nitrophenyl or p-nitrophenyl a-D-glucosides, thus indicating the presence of an a-D-glucosidase activity. For a number of the investigated samples, specific amylolytic activity increased depending on the length of substrates (from p-nitrophenyl maltopentaoside to p-nitrophenyl maltohexaoside); for other IgMs, the opposite dependence was observed. All IgMs studied did not exhibit any other glycoside hydrolase activities toward p-nitrophenyl glycoside substrates.


CONCLUSIONS
Abzyme fractions from different donors demonstrated catalytic heterogeneity in Michaelis-Menten parameters and different modes of action in the hydrolysis of p-nitrophenyl maltooligosaccharides. Enzymatic properties of the IgMs tested varied from human a-amylases. All investigated abzyme samples did not show transglycosylating ability.
Molecular and Radiation Biophysics Division, Petersburg Nuclear Physics Institute of the Russian Academy of Sciences, Gatchina, Russia.
Dina R
DR
Ivanen
Anna A
AA
Kulminskaya
Konstantin A
KA
Shabalin
Luydmila V
LV
Isaeva-Ivanova
Nadezhda A
NA
Ershova
Andrew N
AN
Saveliev
Gregory A
GA
Nevinsky
Kirill N
KN
Neustroev
D000328:Adult; D000681:Amylases / Q000378:metabolism; D017151:Antibodies, Catalytic / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006801:Humans; D007075:Immunoglobulin M / Q000097:blood / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D009103:Multiple Sclerosis / Q000201:enzymology*; D009578:Nitrobenzenes / Q000737:chemistry; D009844:Oligosaccharides / Q000737:chemistry
D017151:Antibodies, Catalytic; D007075:Immunoglobulin M; D009578:Nitrobenzenes; D009844:Oligosaccharides; C075379:4-nitrophenyl maltopentaoside; D000681:Amylases