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Characterization of the self-palmitoylation activity of the transport protein particle component Bet3

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Item Type:Article
Title:Characterization of the self-palmitoylation activity of the transport protein particle component Bet3
Creators Name:Kuemmel, D., Walter, J., Heck, M., Heinemann, U. and Veit, M.
Abstract:Bet3, a transport protein particle component involved in vesicular trafficking, contains a hydrophobic tunnel occupied by a fatty acid linked to cysteine 68. We reported that Bet3 has a unique self-palmitoylating activity. Here we show that mutation of arginine 67 reduced self-palmitoylation of Bet3, but the effect was compensated by increasing the pH. Thus, arginine helps to deprotonate cysteine such that it could function as a nucleophile in the acylation reaction which is supported by the structural analysis of non-acylated Bet3. Using fluorescence spectroscopy we show that long-chain acyl-CoAs bind with micromolar affinity to Bet3, whereas shorter-chain acyl-CoAs do not interact. Mutants with a deleted acylation site or a blocked tunnel bind to Pal-CoA, only the latter with slightly reduced affinity. Bet3 contains three binding sites for Pal-CoA, but their number was reduced to two in the mutant with an obstructed tunnel, indicating that Bet3 contains binding sites on its surface.
Keywords:Golgi, Vesicular Transport, Tethering, TRAPP, Bet3, Palmitoylation, Acyl-CoA
Source:Cellular and Molecular Life Sciences
Page Range:2653-2664
Date:August 2010
Official Publication:https://doi.org/10.1007/s00018-010-0358-y
PubMed:View item in PubMed

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