Epitopes of human brain acetylcholinesterase

The main purpose of the present work was to identify B-cell epitopes on hum an brain acetylcholinesterase (AChE) by the synthetic peptide approach. Fiv e hundred and seventy-four decapeptides comprising amino acids No. n to n+9 (where n denotes the residue number of the 583 amino acids in the primary structure of human brain AChE and is an integer in the range 1-574) were sy nthesized, using the multipin combinatorial chemical synthesis technique, a nd biotinylated. Epitopes of human brain AChE were detected by enzyme-linke d immunosorbent assay (ELISA) and compared with the predicted epitopes of h uman AChE by 'Goldkey' software. Among 574 synthetic decapeptides, 47 decap eptides at 11 antigenic regions showed immunoreactivity with mouse anti-hum an brain AChE polyclonal antibodies. The minimum sequence of epitope was de fined at every antigenic region explored. The locations and sequences of th e farmer ten continuous epitopes at the 11 antigenic regions of the human b rain AChE had been identified as follows: TPVLVWIY (112-119), RTVLVSMNY (14 3-151), LLDQRLALQW (173-182), RRATQLAH (246-253), VFRFSFVPV (294 similar to 302), KDEGSYFLVY (332-341), RVYA (424-427), LMRY (476-479), KAPQWPPY (496- 503), GLRAQACAFW (523-532). The rate of hits of the predicted epitopes from the software came out at 33%. In our work, the epitopes of human AChE have been mapped by purified polyclonal antibody at eleven distinct sites in th e primary structure. (C) 2000 Elsevier Science B.V. All rights reserved.