Lysosome transporter purification and reconstitution identifies Ypq1 pH-gated lysine transport and regulation.

ABSTRACT: Lysosomes achieve their function through numerous transporters that import or export nutrients across their membrane. However, technical challenges in membrane protein overexpression, purification, and reconstitution hinder our understanding of lysosome transporter function. Here, we developed a platform to overexpress and purify the putative lysine transporter Ypq1 using a constitutive overexpression system in protease- and ubiquitination-deficient yeast vacuoles. Using this method, we purified and reconstituted Ypq1 into proteoliposomes and showed lysine transport function, supporting its role as a basic amino acid transporter on the vacuole membrane. We also found that the absence of lysine destabilizes purified Ypq1 and causes it to aggregate, consistent with its propensity to be downr

SUBMITTER: Arines FM 

PROVIDER: S-EPMC10081341 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature