Summary
Based on a structural similarity to the transition state of a propylamine transfer reaction involved in polyamine biosynthesis, Sadenosyl-(5′)-1,8-diamino-3-thiooctane (AdoDATO), the most potent inhibitor of spermidine synthases, was used as a hapten for mice immunization. From immunized mice sera, the IgG fractions were purified by means of affinity (protein A/G) chromatography. Sera and purified polyclonal antibodies from several mice were found to exert spermidine synthase-like activity. Moreover, by means means of hybridoma technology, 19 anti-AdoDATO hybridoma clones have been screened for propylamine transfer activity and at least 6 were found to produce catalytic antibodies. These findings indicate the presence in the sera of active spermidine synthase-like catalytic antibodies. The reported results for the first time evidence the feasibility of preparation of N-alkylating antibodies, widening the biotechnological perspectives of antibodies as biocatalysts.
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Abbreviations
- AdoDATO:
-
S-adenosyl-(5′)-1,8-diaminothiooctane
- TSA:
-
Transition state analog
- decAdoMet:
-
S-adenosyl-(5′)-3-methylthiopropylamine (decarboxylated adenosylmethionine)
- KLH:
-
keyhole limpet hemocyanin
- NMR:
-
nuclear magnetic resonance
- BSA:
-
bovine serum albumine
- WSC:
-
1-ethyl-3-(dimethylaminopropyl)carbodiimide (water-soluble carbodiimide)
- PBS:
-
phosphate buffer saline
- OPD:
-
orthodiphenylenediamine
- TCA:
-
trichloroacetic acid
- SDS-PAGE:
-
sodium duodecylsulphate-polyacrylamide gel electrophoresis
- SN2:
-
bimolecular nucleophilic substitution; abzyme catalytic antibody
- IgG:
-
immunoglobulin G
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Pontoni, G., Lopalco, L., De Maria, S. et al. Studies on propylamine transfer activity in anti-AdoDATO antibodies. Amino Acids 12, 299–308 (1997). https://doi.org/10.1007/BF01373010
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DOI: https://doi.org/10.1007/BF01373010