Interaction of a Golgi-associated kinesin-like protein with Rab6

Science

Volumn 279, Issue 5350, 1998, Pages 580-585

  Echard, Arnaud ^a   Jollivet, Florence ^a   Martinez, Olivier ^a   Lacapère, Jean Jacques ^a   Rousselet, Annie ^a   Janoueix Lerosey, Isabelle ^a   Goud, Bruno ^a  

^a NONE

Author keywords

[No Author keywords available]

Indexed keywords

KINESIN;

ARTICLE; CARBOXY TERMINAL SEQUENCE; ENDOPLASMIC RETICULUM; EUKARYOTIC CELL; GOLGI COMPLEX; INTRACELLULAR TRANSPORT; PRIORITY JOURNAL; PROTEIN ANALYSIS; PROTEIN BINDING; PROTEIN PROTEIN INTERACTION;

ADENOSINE TRIPHOSPHATASES; ALKALINE PHOSPHATASE; AMINO ACID SEQUENCE; BIOLOGICAL TRANSPORT; CARRIER PROTEINS; ENDOPLASMIC RETICULUM; GOLGI APPARATUS; GUANOSINE TRIPHOSPHATE; HELA CELLS; HUMANS; KINESIN; MICROTUBULES; MOLECULAR SEQUENCE DATA; MOLECULAR WEIGHT; RAB GTP-BINDING PROTEINS; RAS PROTEINS;

EUKARYOTA;

EID: 0032559265     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.279.5350.580     Document Type: Article

References (49)

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22. Single-letter abbreviations for the amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; X, any amino acid; and Y, Tyr.
23. + colonies and further analyzed by cotransformation tests and DNA sequencing. pLexA-Rab6, pLexA-Rab6 Q72L, pLexA-Rab6 N126I, pLexA-Rab6 T27N, and pLexA-Rab6 Q22V have been previously described [ I. Janoueix-Lerosey, F. Jollivet, J. Camonis, P. N. Marche, B. Goud J. Biol. Chem. 270, 14801 (1995)], pLexA-Rab6 Q72L,I46E was obtained by cloning from pGEM-1-Rab6 Q72L,I46E (L. Johannes et al., manuscript in preparation).
24. + colonies and further analyzed by cotransformation tests and DNA sequencing. pLexA-Rab6, pLexA-Rab6 Q72L, pLexA-Rab6 N126I, pLexA-Rab6 T27N, and pLexA-Rab6 Q22V have been previously described [ I. Janoueix-Lerosey, F. Jollivet, J. Camonis, P. N. Marche, B. Goud J. Biol. Chem. 270, 14801 (1995)], pLexA-Rab6 Q72L,I46E was obtained by cloning from pGEM-1-Rab6 Q72L,I46E (L. Johannes et al., manuscript in preparation).
25. + colonies and further analyzed by cotransformation tests and DNA sequencing. pLexA-Rab6, pLexA-Rab6 Q72L, pLexA-Rab6 N126I, pLexA-Rab6 T27N, and pLexA-Rab6 Q22V have been previously described [ I. Janoueix-Lerosey, F. Jollivet, J. Camonis, P. N. Marche, B. Goud J. Biol. Chem. 270, 14801 (1995)], pLexA-Rab6 Q72L,I46E was obtained by cloning from pGEM-1-Rab6 Q72L,I46E (L. Johannes et al., manuscript in preparation).
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42. 2, 0.2% bovine serum albumin, 1 mM phenylmethylsulfonyl fluoride, and a mixture of protease inhibitors by incubation at 4°C for 10 min. Supernatants obtained after centrifugation for 10 min at 20,000g were precleared by incubation with protein-G Sepharose for 30 min at 4°C. After preclearing, the supernatants were incubated with 9E10 anti-myc (and in one experiment with affinity-purified anti-Rab6) together with protein-G Sepharose beads for 1 hour at 4°C. The immunoprecipitates were then washed 4 times in the lysis buffer and analyzed by SDS-PAGE and autoradiography.
43. 35S]cysteine (50 μCi/ml).: SEAP present in cell lysates or in the medium was immunoprecipitated with a polyclonal antibody to calf intestine alkaline phosphatase (Rockland).
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49. We thank M. Bornens, M.-H. Cuif, B. Hoflack, L. Johannes, G. Langsley, J. de Mey, M. Kress, J.-L. Rigaud, J. Salamero, and C. Saudrais for critical reading of this manuscript and helpful discussions; M. Bornens (Institut Curie, Paris), E. Berger (Zurich, Switzerland), M. Zerial [European Molecular Biology Laboratory (EMBL), Heidelberg, Germany], and P. Chavrier (Centre d'Immunologie de Marseille-Luminy, Marseille, France) for the gifts of CTR 433 monoclonal antibody, affinity-purified polyolonal antibody to human β-1,4-galactosyltransferase, pLexA-Rab5 Q79L, and pLexA-Rab7 Q67L, respectively; and G. Bordenave for help in rabbit immunization. Supported in part by grants from the Association de la Recherche Contre le Cancer, the Fondation de la Recherche Médicale, the European Community, and the Human Frontier Science Program.