L-proline Transport Systems of Starfish Pyloric Caeca

Purified brush-border membrane vesicles (BBMV) of starfish [Pycnopodia helianthoides (Brandt)] pyloric caecal epithelium were prepared by magnesium precipitation in order to characterize the possible role of this organ in amino acid transport. L-[H]proline uptake by these vesicles was Na-dependent and greater at pH7.5 than at pH5.5. L-Pipecolate was a competitive inhibitor of L-proline influx into these BBMV, exhibiting a Kx value of O.fGmmoir . The amino acid inhibitors, L-pipecolate, L-alanine and L-leucine were used as test substrates to block L-prohne influx by the IMINO, NBB and L transport systems, respectively, in order to estimate the contribution of each process to total L-prohne entry into pyloric caecal cells. The carrier-mediated transport constants for L-prohne transfer by these three systems were: /Ct=0.18mmoll~ 1 (IMINO), 0 13mmol I" (NBB) and ( U l m m o i r 1 (L); /m a x= 1310 pmol nig" 1 protein 30 s" (IMINO), 360 pmolmg" protein 30 s 1 (NBB) and 470pmolmg"protein30s" (L). L-Proline influxes through both the IMINO and NBB systems were sigmoidal functions of the external [Na], while transfer by the L system was Naindependent. Multiple sodium ions (e.g. 2 or 3 Na/L-proline) appear to be associated with L-proline transport by both Na-dependent transport systems, but the nature of this association (i.e. activation or energization) is unclear. Results suggest that starfish pyloric caecal epithelium possesses a similar array of L-proline transport proteins to those found in similar cell types of mammalian intestine or kidney, providing tentative support for an absorptive function for this organ.