Title

Purification of the Golgi adenosine 3'-phosphate 5'-phosphosulfate transporter, a homodimer within the membrane

UMMS Affiliation

Department of Biochemistry and Molecular Pharmacology

Date

10-25-1994

Document Type

Article

Subjects

Affinity Labels; Animals; Binding, Competitive; Carrier Proteins; Chromatography, Affinity; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Dose-Response Relationship, Radiation; Electrophoresis, Polyacrylamide Gel; Golgi Apparatus; Intracellular Membranes; Kinetics; Liposomes; Liver; Molecular Weight; Phosphoadenosine Phosphosulfate; Proteolipids; Rats

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

Sulfation of proteoglycans, secretory and membrane proteins, and glycolipids occurs in the lumen of the Golgi apparatus. Adenosine 3'-phosphate 5'-phosphosulfate (PAPS), the sulfate donor in these reactions, must be transported from the cytosol, its site of synthesis, into the lumen of the Golgi apparatus. We have identified and purified to apparent homogeneity the rat liver Golgi membrane PAPS transporter by a combination of conventional and affinity chromatography as well as photoaffinity radiolabeling with adenosine 3',5'-bisphosphate, a competitive inhibitor of PAPS transport. The transporter, a 75-kDa protein, was purified 70,000-fold over homogenate (6% yield) and transported PAPS into phosphatidylcholine liposomes selectively and in a saturable manner (apparent Km of 1.7 microM). Radiation target-inactivation analyses of the transport activity in rat liver Golgi vesicles, together with the above described biochemical approaches, demonstrate that the PAPS transporter within the Golgi membrane is a homodimer.

Rights and Permissions

Citation: Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10707-11.

Related Resources

Link to Article in PubMed

Journal Title

Proceedings of the National Academy of Sciences of the United States of America

PubMed ID

7938015