Solubilization and reconstitution characteristics of hepatic system A-mediated amino acid transport.

摘要: In the liver, System A-mediated uptake of neutral amino acids may play a key role in metabolic control. Knowing properties solubilized and reconstituted A activity is important for future studies on purification carrier protein. Solubilization by combination 2.5% cholate 4 M urea resulted greater than 85% extraction activity. Previous removal easily extracted plasma membrane proteins with 1% alone followed solubilization transporter cholate/urea 2-fold enrichment transport Based observation that protein aggregates presence low detergent concentrations, selective polyethylene glycol precipitation procedure was developed resulting recovery more 70% initial less 10% total concomitant 10-fold achieved. The precipitated could be resuspended buffer containing Triton X-100, asolectin, glycerol. Transporter this stable up to 5 days when maintained at -20 degrees C or 2 C. general applicability devised reconstitution illustrated Systems N Gly proteoliposomes specific activities those native vesicles.