vps29 is not an active metallo-phosphatase but is a rigid scaffold required for retromer interaction with accessory proteinsvps29不是积极metallo-phosphatase而是是一个刚性支架retromer与辅助交互所需的蛋白质.pdfVIP

VPS29 Is Not an Active Metallo-Phosphatase but Is a Rigid Scaffold Required for Retromer Interaction with Accessory Proteins 1 2 1 2 3¤ James D. Swarbrick , Daniel J. Shaw , Sandeep Chhabra , Rajesh Ghai , Eugene Valkov , Suzanne J. 2 4 2 Norwood , Matthew N. J. Seaman , Brett M. Collins * 1 Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia, 2 Institute for Molecular Bioscience, The University of Queensland, St. Lucia, Queensland, Australia, 3 School of Chemistry and Molecular Bioscience, The University of Queensland, St. Lucia, Queensland, Australia, 4 Department of Clinical Biochemistry, Cambridge Institute for Medical Research, Cambridge University, Cambridge, United Kingdom Abstract VPS29 is a key component of the cargo-binding core complex of retromer, a protein assembly with diverse roles in transport of receptors within the endosomal system. VPS29 has a fold related to metal-binding phosphatases and mediates interactions between retromer and other regulatory proteins. In this study we examine the functional interactions of mammalian VPS29, using X-ray crystallography and NMR spectroscopy. We find that although VPS29 can coordinate metal ions Mn2+ and Zn2+ in both the putative active site and at other locations, the affinity for metals is low, and lack of activity in phosphatase assays using a putative peptide substrate support the conclusion that VPS29 is not a functional metalloenzyme. There is evidence that structural elements of VPS29 critical