| Abstract: | Antibodies against the alpha and beta subunits of phenylalanyl-tRNA synthetase were fractionated by ion exchange chromatography into different classes and then digested with papain to yield the respective Fab fragments. The preparations obtained were used to investigate (i) whether the alpha and beta polypeptides share any common antigenic determinants and (ii) whether immunological methods are able to resolve the catalytic function of the subunits of this enzyme (or principally of oligomeric enzymes). As to the first problem, immunodiffusion and complement fixation experiments showed that there is no immunological relatedness between the subunits which argues against the existence of sequence homoligies. As to the second question investigated, it was found that any binding of immunoglobulins of Fab fragments to the alpha or the the beta subunit affects enzyme activity either in the direction of activation or inhibition. These results therefore show that the immunological approach is not appropriate for resolving subunit-specific funcitons, possibly as a consequence of conformational changes induced in the enzyme by the binding of the immunoglobulins of Fab fragments. |
