Pik1 online dating

We now report the x-ray crystal structure of human GOLPH3 to a 2.9-Å resolution.Crystals of GOLPH3 lacking the first 51 amino acids (GOLPH3Δ51) grew from a solution containing ∼0.9 M ammonium sulfate and contain bound sulfate ions that suggest the location of a Ptd Ins4 and Fig.Dioleoylphosphatidylcholine (DOPC) vesicles with or without 3% (mol/mol) Ptd Ins3 value of 2.6 ± 0.2 µM, which is consistent with the more robust targeting of GFP-GOLPH3 to the Golgi compared with GFP-Vps74.

In yeast, steady-state Golgi localization of multiple mannosyltransferases requires recognition of their cytosolic domains by the peripheral Golgi membrane protein Vps74, an orthologue of human GOLPH3/GPP34/GMx33/MIDAS (mitochondrial DNA absence sensitive factor).

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We now report that recruitment of yeast Vps74 and human GOLPH3 to the yeast Golgi apparatus requires signaling by the Golgi-localized Ptd Ins 4-kinase Pik1 and specific recognition of Ptd Ins 4-phosphate (Ptd Ins4) in a conserved binding pocket on the surface of Vps74 and GOLPH3.

Golgi mannosyltransferases that depend on Vps74 for Golgi localization are not retained in the Golgi in mutant cells that express a form of Vps74 that does not bind Ptd Ins4To identify factors required to recruit Vps74 to the Golgi, we used fluorescence microscopy to screen a collection of yeast mutants for defects in Golgi localization of a GFP-Vps74 fusion protein.

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Mutations in lead to cell death by impairing secretion at multiple points within the secretory pathway, possibly via misregulation of Arf GTPases (Garcia-Bustos et al., 1994; Hama et al., 1999; Walch-Solimena and Novick, 1999; Audhya et al., 2000; Sciorra et al., 2005; Strahl et al., 2005).

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