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  • Title: Comparison of the effects of certain thiol reagents on alanine transport in plasma membrane vesicles from rat liver and their use in identifying the alanine carrier.
    Author: Hayes MR, McGivan JD.
    Journal: Biochem J; 1983 Aug 15; 214(2):489-95. PubMed ID: 6615478.
    Abstract:
    The Na+-dependent uptake of alanine into plasma membrane vesicles from rat liver was inhibited by N-ethylmaleimide (NEM) and by mersalyl. NEM did not inhibit alanine-independent Na+ uptake and the inhibition of alanine transport by NEM was protected by pre-incubation with an excess of substrate. It was therefore concluded that NEM acted by binding to the alanine carrier. A protein of Mr 20 000 was found to bind NEM with a concentration dependence parallel to the NEM inhibition of alanine transport. The inhibition of binding of [3H]NEM to this protein by mersalyl had a concentration dependence similar to that of the inhibition of transport by mersalyl. Preincubation with L-alanine, but not with D-alanine, led to protection of the Mr 20 000 protein from binding NEM. It is concluded that this protein is an essential component of the alanine transport system.
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