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EMBO J. Jun 15, 1998; 17(12): 3258–3268.
PMCID: PMC1170664

Mapping the interaction between GRASP65 and GM130, components of a protein complex involved in the stacking of Golgi cisternae.


The nature of the complex containing GRASP65, a membrane protein involved in establishing the stacked structure of the Golgi apparatus, and GM130, a putative Golgi matrix protein and vesicle docking receptor, was investigated. Gel filtration revealed that GRASP65 and GM130 interact in detergent extracts of Golgi membranes under both interphase and mitotic conditions, and that this complex can bind to the vesicle docking protein p115. Using in vitro translation and site-directed mutagenesis in conjunction with immunoprecipitation, the binding site for GRASP65 on GM130 was mapped to the sequence xxNDxxxIMVI-COOH at the C-terminus of GM130, a region known to be required for its localization to the Golgi apparatus. The same approach was used to show that the binding site for GM130 on GRASP65 maps to amino acids 189-201, a region conserved in the mammalian and yeast proteins and reminiscent of PDZ domains. Using green fluorescent protein (GFP)-tagged reporter constructs, it was shown that one essential function of the interaction between GRASP65 and GM130 is in the correct targeting of the two proteins to the Golgi apparatus.

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Selected References

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  • Ackers GK. Analytical gel chromatography of proteins. Adv Protein Chem. 1970;24:343–446. [PubMed]
  • Amos WB, Grimstone AV. Intercisternal material in the golgi body of Trichomonas. J Cell Biol. 1968 Aug;38(2):466–471. [PMC free article] [PubMed]
  • Barr FA, Puype M, Vandekerckhove J, Warren G. GRASP65, a protein involved in the stacking of Golgi cisternae. Cell. 1997 Oct 17;91(2):253–262. [PubMed]
  • Bhatnagar RS, Gordon JI. Understanding covalent modifications of proteins by lipids: where cell biology and biophysics mingle. Trends Cell Biol. 1997 Jan;7(1):14–20. [PubMed]
  • Burke B, Griffiths G, Reggio H, Louvard D, Warren G. A monoclonal antibody against a 135-K Golgi membrane protein. EMBO J. 1982;1(12):1621–1628. [PMC free article] [PubMed]
  • Cluett EB, Brown WJ. Adhesion of Golgi cisternae by proteinaceous interactions: intercisternal bridges as putative adhesive structures. J Cell Sci. 1992 Nov;103(Pt 3):773–784. [PubMed]
  • Cowburn D. Adaptors and integrators. Structure. 1996 Sep 15;4(9):1005–1008. [PubMed]
  • Devereux J, Haeberli P, Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. [PMC free article] [PubMed]
  • Franke WW, Kartenbeck J, Krien S, VanderWoude WJ, Scheer U, Morré DJ. Inter- and intracisternal elements of the Golgi apparatus. A system of membrane-to-membrane cross-links. Z Zellforsch Mikrosk Anat. 1972;132(3):365–380. [PubMed]
  • Fritzler MJ, Hamel JC, Ochs RL, Chan EK. Molecular characterization of two human autoantigens: unique cDNAs encoding 95- and 160-kD proteins of a putative family in the Golgi complex. J Exp Med. 1993 Jul 1;178(1):49–62. [PMC free article] [PubMed]
  • Fritzler MJ, Lung CC, Hamel JC, Griffith KJ, Chan EK. Molecular characterization of Golgin-245, a novel Golgi complex protein containing a granin signature. J Biol Chem. 1995 Dec 29;270(52):31262–31268. [PubMed]
  • Johnson DR, Bhatnagar RS, Knoll LJ, Gordon JI. Genetic and biochemical studies of protein N-myristoylation. Annu Rev Biochem. 1994;63:869–914. [PubMed]
  • Kooy J, Toh BH, Pettitt JM, Erlich R, Gleeson PA. Human autoantibodies as reagents to conserved Golgi components. Characterization of a peripheral, 230-kDa compartment-specific Golgi protein. J Biol Chem. 1992 Oct 5;267(28):20255–20263. [PubMed]
  • Kornau HC, Schenker LT, Kennedy MB, Seeburg PH. Domain interaction between NMDA receptor subunits and the postsynaptic density protein PSD-95. Science. 1995 Sep 22;269(5231):1737–1740. [PubMed]
  • Levine TP, Rabouille C, Kieckbusch RH, Warren G. Binding of the vesicle docking protein p115 to Golgi membranes is inhibited under mitotic conditions. J Biol Chem. 1996 Jul 19;271(29):17304–17311. [PubMed]
  • Linstedt AD, Hauri HP. Giantin, a novel conserved Golgi membrane protein containing a cytoplasmic domain of at least 350 kDa. Mol Biol Cell. 1993 Jul;4(7):679–693. [PMC free article] [PubMed]
  • Lucocq JM, Pryde JG, Berger EG, Warren G. A mitotic form of the Golgi apparatus in HeLa cells. J Cell Biol. 1987 Apr;104(4):865–874. [PMC free article] [PubMed]
  • Misteli T, Warren G. COP-coated vesicles are involved in the mitotic fragmentation of Golgi stacks in a cell-free system. J Cell Biol. 1994 Apr;125(2):269–282. [PMC free article] [PubMed]
  • Misumi Y, Sohda M, Yano A, Fujiwara T, Ikehara Y. Molecular characterization of GCP170, a 170-kDa protein associated with the cytoplasmic face of the Golgi membrane. J Biol Chem. 1997 Sep 19;272(38):23851–23858. [PubMed]
  • Moremen KW, Touster O, Robbins PW. Novel purification of the catalytic domain of Golgi alpha-mannosidase II. Characterization and comparison with the intact enzyme. J Biol Chem. 1991 Sep 5;266(25):16876–16885. [PubMed]
  • Nakamura N, Rabouille C, Watson R, Nilsson T, Hui N, Slusarewicz P, Kreis TE, Warren G. Characterization of a cis-Golgi matrix protein, GM130. J Cell Biol. 1995 Dec;131(6 Pt 2):1715–1726. [PMC free article] [PubMed]
  • Nakamura N, Lowe M, Levine TP, Rabouille C, Warren G. The vesicle docking protein p115 binds GM130, a cis-Golgi matrix protein, in a mitotically regulated manner. Cell. 1997 May 2;89(3):445–455. [PubMed]
  • Ponting CP. Evidence for PDZ domains in bacteria, yeast, and plants. Protein Sci. 1997 Feb;6(2):464–468. [PMC free article] [PubMed]
  • Ponting CP, Phillips C, Davies KE, Blake DJ. PDZ domains: targeting signalling molecules to sub-membranous sites. Bioessays. 1997 Jun;19(6):469–479. [PubMed]
  • Rambourg A, Clermont Y. Three-dimensional electron microscopy: structure of the Golgi apparatus. Eur J Cell Biol. 1990 Apr;51(2):189–200. [PubMed]
  • Sapperstein SK, Walter DM, Grosvenor AR, Heuser JE, Waters MG. p115 is a general vesicular transport factor related to the yeast endoplasmic reticulum to Golgi transport factor Uso1p. Proc Natl Acad Sci U S A. 1995 Jan 17;92(2):522–526. [PMC free article] [PubMed]
  • Saras J, Heldin CH. PDZ domains bind carboxy-terminal sequences of target proteins. Trends Biochem Sci. 1996 Dec;21(12):455–458. [PubMed]
  • Seelig HP, Schranz P, Schröter H, Wiemann C, Renz M. Macrogolgin--a new 376 kD Golgi complex outer membrane protein as target of antibodies in patients with rheumatic diseases and HIV infections. J Autoimmun. 1994 Feb;7(1):67–91. [PubMed]
  • Slusarewicz P, Nilsson T, Hui N, Watson R, Warren G. Isolation of a matrix that binds medial Golgi enzymes. J Cell Biol. 1994 Feb;124(4):405–413. [PMC free article] [PubMed]
  • Songyang Z, Fanning AS, Fu C, Xu J, Marfatia SM, Chishti AH, Crompton A, Chan AC, Anderson JM, Cantley LC. Recognition of unique carboxyl-terminal motifs by distinct PDZ domains. Science. 1997 Jan 3;275(5296):73–77. [PubMed]
  • Thyberg J, Moskalewski S. Microtubules and the organization of the Golgi complex. Exp Cell Res. 1985 Jul;159(1):1–16. [PubMed]
  • Waters MG, Clary DO, Rothman JE. A novel 115-kD peripheral membrane protein is required for intercisternal transport in the Golgi stack. J Cell Biol. 1992 Sep;118(5):1015–1026. [PMC free article] [PubMed]

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