HYDROLYSIS OF TRANSFORMING GROWTH FACTOR-ALPHA BY CELL-SURFACE PEPTIDASES INVITRO

Human transforming growth factor-alpha (h-TGF-alpha), a 50-amino acid residue peptide, was incubated with some purified cell-surface peptidases and with renal microvillar membranes prepared from pig and rat. Hydrolysis was monitored by h.p.l.c. and activity by a biological assay. Prolonged incubation with relatively large amounts of endopeptidase-24.11, aminopeptidase N and peptidyl dipeptidase A (angiotensin-converting enzyme) caused no observable hydrolysis and no detectable loss of biological activity. Incubation with pig renal microvilli also failed to degrade the peptide. In contrast. rat renal microvilli readily degraded h-TGF-alpha, as did endopeptidase-2, which is located in rat renal and intestinal brush borders, but is absent from pig kidneys. This enzyme degraded about 30 nmol of h-TGF-alpha/h per mg of protein. The physiological significance of these results is discussed.