THE EFFECT OF A TRYPSIN INHIBITOR, TRASYLOL, ON THE COURSE OF BILE AND TRYPSIN-INDUCED PANCREATITIS IN DOGS.
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A unique trypsinogen complementary DNA has been isolated from an Atlantic cod (Gadus morhua) cDNA library. Its predicted amino acid sequence contains 249 residues with a putative polypeptide of 227 residues. The distinctive features of this polypeptide, referred to as trypsin Y, are its low number of hydrogen-bond-forming residues, high content of Met and Pro residues, and lack of one conserved disulfide bond. Alignments show that cod trypsinogen Y has only approximately 45% identity to the two Atlantic cod trypsinogens I and X and most vertebrate trypsinogens. However, it has more than 70% identity to three other fish trypsinogens from two Pleuronectes and an Antarctic Notothenia species. These four trypsinogens share some unique characteristics and form a novel group, here referred to as group III.
Complementary DNAs encoding precursors of the three heavy chains (HC1, HC2, HC3) of the inter-alpha-trypsin inhibitor in Syrian hamster liver were sequenced. The deduced amino acid sequence of the HC1 precursor was 87, 82, and 79% identical with those of the HC1 precursors from mouse, man and pig, respectively. The HC2 and HC3 precursors showed similar degrees of sequence identity with the corresponding human and mouse HC precursors. When the hamster HC1 precursor was compared with its own HC2 and HC3 precursors, however, even the most highly conserved segment consisting of 565 amino acid residues, i.e., about 2/3 of the whole molecule, showed only about 35 and 65% sequence identity, respectively. Essentially the same results were obtained on the intra-species comparisons of three subfamilies in man and mouse. Thus, the interspecies conservation of a given HC subfamily is much greater than the similarity between the three different HC subfamilies within a given species. These results suggest that (i) higher vertebrates possess three HC genes which have been evolving independently of each other under purifying selection; (ii) the diversification of the three HC subfamilies, for which the middle regions of the molecules were mainly responsible, occurred before eutherian radiation; and (iii) each HC subfamily may have unique function(s), although at present virtually nothing is known about the functional differences between the three HC subfamilies.
Distinguished lymphocytes stimulation was observed in vitro with one out of four patients received Trasylol therapy. The peak response of DNA synthesis was shown to occur at 100 mug/tube of basic pancreatic trypsin inhibitor (BPTI) itself, about 6 days after addition of BPTI. Reactivity (3H-thymidine incorporation) of lymphocytes of patients after the therapy, to specific antigen, ie., purified protein derivative (PPD) or Bordetella pertussis organisms in vitro, was markedly depressed. On the other hand, that to non-specific mitogen, phytohaemagglutinin-p of pokeweed mitogen, was a little reduced, or rather enhanced after the therapy. Conversly, lymphocytes of an originally tuberculin negative patient were lead to be stimulated with PPD in vitro, after skin test with PPD and Trasylol therapy. These various immune responses including drug-induced exanthemas of patients received the therapy, are discussed in this report, in comparison with immune responses of guinea pigs sensitized with BPTI, immune responses of animals in microorganism infections and the contrary immunological effects of Trasylol already reported.
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