Synthetic peptides of the Rab effector domain inhibit vesicular transport through the secretory pathway.

Plutner, Helen; Schwaninger, Ruth; Pind, Steven; Balch, William E.

doi:10.1002/j.1460-2075.1990.tb07412.x

Cited by 134 publications

(83 citation statements)

References 67 publications

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“…Peptides corresponding to the effector domain of rab3, ras and rabl were synthesized as prevtously described [16]. GTPyS and GDP/JS were purchased from Boehrmger Mannheim.…”

Section: Methodsmentioning

confidence: 99%

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Stimulation of insulin release from permeabilized HIT‐T15 cells by a synthetic peptide corresponding to the effector domain of the small GTP‐binding protein rab3

Regazzi

Balch

et al. 1993

FEBS Letters

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A synthetic peptide (rab3AL) correspondmg to the erector domam of rab3, a small GTP-bmdmg protein, stimulated basal and potentiated Ca"-as well as GTPyS-evoked insuhn secretion about Z-fold from streptolysin-0 permeablhzed HIT cells. This effect was specific, since the analogous peptides of ras or rabl did not affect the exocytotic event. The more than additlre effect of rab3AL on Ca" or GTPyS stimulation indicates a distinct mode of action of the peptide. The partial loss of cytosohc proteins from permeabilized ceils was accompanied by a faster run-doun of the secretory response to Ca" than the one to GTP@. The persistent effect of rab3AL under these conditions points to a inembrane locahzation of Its target. These results suggest that rab3 and its effector are involved m the regulation of insulin secretion.

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“…Peptides corresponding to the effector domain of rab3, ras and rabl were synthesized as prevtously described [16]. GTPyS and GDP/JS were purchased from Boehrmger Mannheim.…”

Section: Methodsmentioning

confidence: 99%

“…A synthetic peptide corresponding to the putative effector domain of this SMG, except for the exchange of two amino acids (denoted rab3AL). was shown to affect intracellular vesicle transport [16].…”

Section: Introductionmentioning

confidence: 99%

Stimulation of insulin release from permeabilized HIT‐T15 cells by a synthetic peptide corresponding to the effector domain of the small GTP‐binding protein rab3

Regazzi

Balch

et al. 1993

FEBS Letters

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“…By analogy with ras, the effector domain is believed to interact with a GAP protein, thereby stimulating the low endogenous GTPase activity of rab proteins. Effector domain peptides have, in fact, been shown to inhibit transport in the biosynthetic pathway (Plutner et al, 1990). Proteins have been identified that can escort rab proteins in the cytosol (GDI), stimulate GTP hydrolysis or facilitate GDP/GTP exchange (Huang et al, 1990;Sasaki et al, 1990;West et al, 1990;Burstein et al, 1991;Tan et al, 1991;Burstein and Macara, 1992).…”

Section: Introductionmentioning

confidence: 99%

The N-terminal domain of a rab protein is involved in membrane-membrane recognition and/or fusion.

et al. 1994

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Proteins of the YPT1/SEC4/rab family are well documented to be involved in the regulation of membrane transport. We have previously reported that rab5 regulates endosome-endosome recognition and/or fusion in vitro. Here, we show that this process depends on the rab5 N-terminal domain. Treatment of early endosomal membranes at a low trypsin concentration essentially abolished fusion and cleaved rab5 to a 1 kDa smaller polypeptide. Two-dimensional gel analysis suggested that rab5 is one of the few, if not the only, polypeptides cleaved by trypsin under these conditions. Whereas endosome fusion could be stimulated by cytosol prepared from cells overexpressing rab5 (and thus containing high amounts of the protein), this stimulation was abolished by trypsin-treatment of the cytosol. Trypsin-treated cytosol prepared from mock-transfected cells, which contains very low amounts of rab5, showed no inhibitory activity indicating that rab5 is the target of trypsin in these experiments. Purified rab5 prepared after expression in Escherichia coli was treated with trypsin, which cleaved the protein at the N-terminus. A synthetic peptide of rab5 N-terminal domain inhibited endosome fusion in our cell-free assay. A version of the same peptide truncated at the N-terminus or a peptide of rab3 N-terminal domain were without effects. Altogether, these observations suggest that the N-terminal domain of rab5 is involved in the process of early endosome recognition and/or fusion, presumably because it interacts with another component of the transport machinery.

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“…The peptide nomenclature used follows that of Plutner et al [17]. The numbering refers to the homologous amino acid positions in Ha-ras.…”

Section: Methodsmentioning

confidence: 99%

Inhibition of pituitary hormone exocytosis by a synthetic peptide related to the rab effector domain

et al. 1993

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GTP-binding proteins of the rab family are believed to function at several steps in intracellular vesicular transport. We examined the effects of a rab-related peptide in permeabilized pituitary cells, in which exocytosis can be triggered by distinct Ca2+-dependent or Ca:+-independent pathways. We report that a synthetic peptide of 18 amino acids related to the rab effector domain, rab3AL(30~,7) inhibited luteinizing hormone (LH) and growth hormone (GH) exocytosis triggered by either pathway. Ca:+-stimulated LH and GH release were inhibited by more than 80% and 50%, respectively, by 100 ,uM peptide. The peptide (100/~M) also inhibited LH and GH exocytosis stimulated by phorbol myristate acetate plus cAMP by more than 45% and 80%, respectively. The effect was sequence-specific since a second peptide, lacking the first 3 amino acids but otherwise identical failed to inhibit exocytosis. These results suggest that a protein of the rab family is involved in regulated pituitary hormone exocytosis, and they identify 3 amino acids of the putative rab effector domain which may be functionally important in exocytosis.

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Synthetic peptides of the Rab effector domain inhibit vesicular transport through the secretory pathway.

Cited by 134 publications

References 67 publications

Stimulation of insulin release from permeabilized HIT‐T15 cells by a synthetic peptide corresponding to the effector domain of the small GTP‐binding protein rab3

Stimulation of insulin release from permeabilized HIT‐T15 cells by a synthetic peptide corresponding to the effector domain of the small GTP‐binding protein rab3

The N-terminal domain of a rab protein is involved in membrane-membrane recognition and/or fusion.

Inhibition of pituitary hormone exocytosis by a synthetic peptide related to the rab effector domain

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