Golgins tethering

J., Satch, A., Pelletier, L., and Warren, G., Golgin tethers define subpopulations of COPI vesicles. Science 307,1095-1098,2005 Short, B Haas, A., and Barr, F.A., Golgins and GTPases, giving identity and structure to the Golgi apparatus, Biochim. Biophys. Acta 1744, 383-395, 2005 Satoh, A., Beard, M., and Warren, G., Preparation and characterization of recombinant golgin tethers. Methods Enzymol. 404, 279-296, 2005. 

Note that uncoating the COPI vesicles is required for binding to golgin tethers (or Golgi membranes, see below) since binding was not observed when an ArflQVlL mutant was used for preparation of COPI vesicles (Malsam et al, 2005). 

Malsam, J., Satoh, A., Pelletier, L., and Warren, G. (2005). Golgin tethers define subpopulations of COPI vesicles. Science 307, 1095-1098. 

PREPARATION AND CHARACTERIZATION OE RECOMBINANT GOLGIN TETHERS 295 Acknowledgment... 

TETHERING ASSAYS FOR COPI VESICLES MEDIATED BY GOLGINS... 

In Vitro COPI Vesicle Tethering Assay Using Purified Golgins... 

In Vitro Tethering of COPI Vesicles to Golgin-Coated Glass Slides... 

Fig. 1. Tethering of COPI vesicles to CASP-coated slides. (A-E) The vesicle-containing supernatant was incubated for 30 min at 4° on glass slides precoated with (A, D, E) recombinant CASP, (B) 1% BSA and STI, or (C) recombinant EEAl in the absence (A, B, C) or presence of (D) soluble CASP or (E) soluble golgin-84. Glass slides were embedded in Epon resin and processed for conventional EM. (F) Input membranes. (G) Quantitation of the results in (A-E) presented as the mean of the number of the bound vesicles per fim S. D.(n=5). (Reprinted with permission from Science.)...

Golgins are minor components of Golgi membranes so that preparation of sufficient amounts for biochemical analysis is difficult. Here we describe the preparation of recombinant forms of those golgins that constitute the tethering complexes just described. 

Shorter, J., Beard, M. B., Seemann, J., Dirac-Svejstrup, A. B., and Warren, G. (2002). Sequential tethering of Golgins and catalysis of SNAREpin assembly by the vesicle-tethering protein pll5. J. Cell Biol 157, 45-62. 

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