Effect of phospholipase A on active transport of amino acids with membrane vesicles of Mycobacterium phlei.

Active transport of proline remained unaffected in phospholipase A-treated electron transport particles from Mycobacterium phlei. However, the steady state level of proline was reduced 50 to 60% in phospholipase A-treated depleted electron transport particles that were devoid of membrane-bound coupling factor-latent ATPase activity. The decrease in the uptake of proline in the phospholipase A-treated depleted electron transport particles was not due to a change in the apparent K-m for proline, but it was related to the amount of phospholipid cleaved from the membranes. Restoration in the level of proline transport in phospholipase A-treated depleted electron transport particles was achieved by reconstituting these vesicles with diphosphatidylglycerol and phosphatidylethanolamine liposomes. Diphosphatidylglycerol was found to be most effective in the restoration of proline uptake. In contrast to the effect of phospholipase A treatment on proline transport, similar treatement of the electron transport particles or depleted electron transport particles failed to inhibit the active transport of either glutamine or glutamic acid. Studies with phospholipase A-treated membrane vesicles confirmed earlier findings that a proton gradient is not required for active transport of amino acids.