Human small intestinal angiotensin-converting enzyme: intracellular transport, secretion and glycosylation

Human intestinal angiotensin-converting enzyme (ACE) exists in the brush-border membrane as a monomeric protein of apparent molecular mass 184 kDa. It is associated with the membrane via a hydrophobic segment and has a transmembrane orientation [Naim (1992) Biochem. J. 286, 451-457]. In addition to the membrane-bound form (ACEm), hydrophilic forms of ACE (ACEsec) can be identified in biosynthetically labelled intestinal cells. Thus the culture medium of biosynthetically labelled human biopsy samples contains an ACE molecule which has an apparent molecular mass similar to that of its membrane-bound counterpart. The secreted ACEsec forms follow a precursor/product relationship with the mature ACE molecule. The effect of the monomeric structure of ACE in its intracellular transport and secretion was investigated by pulse-chase experiments on human biopsy samples labelled with [35S]methionine. The results reveal 2-3-fold slower transport of ACE from the endoplasmic reticulum (ER) to the Golgi as compared with the homodimeric proteins dipeptidylpeptidase IV and aminopeptidase N. Further, the transport kinetics of ACE are comparable with those of human sucrase-isomaltase and human maltase-glucoamylase, two brush-border disaccharidases that do not form homodimers in the ER of human small-intestinal cells. These findings strongly suggest that homodimerization of brush-border proteins may influence the rate of transport of these proteins from the ER to the Golgi. The effect of glycosylation on the transport and secretion of ACE was investigated by utilizing several inhibitors of glycan processing. Here, secretion of ACE molecules continued to take place, albeit to a considerably lesser extent. In fact, approx. 2-fold less ACE molecules were secreted in the presence of inhibitors of ER glucosidases I and II and cis-Golgi mannosidase-I, suggesting that carbohydrate processing is important in the attainment of a transport-competent conformation.