Regioselective enzymatic acylation of N-acetylhexosamines

Simerská, Pavla, Pišvejcová, Andrea, Kuzma, Marek, Sedmera, Petr, Křen, Vladimır, Nicotra, Silvia and Riva, Sergio (2004) Regioselective enzymatic acylation of N-acetylhexosamines. Journal of Molecular Catalysis B: Enzymatic, 29 1-6: 219-225. doi:10.1016/j.molcatb.2003.10.018


Author Simerská, Pavla
Pišvejcová, Andrea
Kuzma, Marek
Sedmera, Petr
Křen, Vladimır
Nicotra, Silvia
Riva, Sergio
Title Regioselective enzymatic acylation of N-acetylhexosamines
Journal name Journal of Molecular Catalysis B: Enzymatic   Check publisher's open access policy
ISSN 1873-3158
Publication date 2004-01-01
Sub-type Article (original research)
DOI 10.1016/j.molcatb.2003.10.018
Open Access Status DOI
Volume 29
Issue 1-6
Start page 219
End page 225
Total pages 7
Place of publication Amsterdam, Netherlands
Publisher Elsevier
Language eng
Abstract S-acylation/deacylation cycles and vesicular transport are critical for an adequate subcellular distribution of S-acylated Ras proteins. H-Ras is dually acylated on cysteines 181 and 184, but it is unknown how these residues individually contribute to H-Ras trafficking. In this study, we characterized the acylation and deacylation rates and membrane trafficking of monoacylated H-Ras mutants to analyze their contributions to H-Ras plasma membrane and endomembrane distribution. We demonstrated that dually acylated H-Ras interacts with acyl-protein thioesterases (APTs) 1 and 2 at the plasma membrane. Moreover, single-acylation mutants of H-Ras differed not only in their subcellular distribution, where both proteins localized to different extents at both the Golgi complex and plasma membrane, but also in their deacylation rates, which we showed to be due to different sensitivities to APT1 and APT2. Fluorescence photobleaching and photoactivation experiments also revealed that 1) although S-acylated, single-acylation mutants are incorporated with different efficiencies into Golgi complex to plasma membrane vesicular carriers, and 2) the different deacylation rates of single-acylated H-Ras influence differentially its overall exchange between different compartments by nonvesicular transport. Taken together, our results show that individual S-acylation sites provide singular information about H-Ras subcellular distribution that is required for GTPase signaling.
Formatted abstract
A careful choice of the reaction conditions (solvent, enzyme, acylating agent) allowed an efficient regioselective acylation of N-acetylhexosamines. 6-O-Acyl derivatives of 2-acetamido-2-deoxy-D-glucopyranose (GlcNAc), 2-acetamido-2-deoxy-D-galactopyranose (GalNAc) and 2-acetamido-2-deoxy-D- mannopyranose (ManNAc) have been isolated from regioselective esterifications catalysed by the protease subtilisin in CH3CN-DMSO 8:2 in good yields.
Keyword Enzymatic acylation
N-acetylhexosamines
Regioselectivity
Subtilisin
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Chemistry and Molecular Biosciences
 
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Created: Wed, 28 Sep 2011, 02:46:55 EST by Dr Pavla Simerska on behalf of School of Chemistry & Molecular Biosciences