De-N-acetylation of N-acetylglucosaminyl-phosphatidylinositol (GlcNAc-PI) is the second step of glycosylphosphatidylinositol (GPI) membrane anchor biosynthesis in eukaryotes. This step is a prerequisite for the subsequent mannosylation of glucosaminyl-phosphatidylinositol (GlcN-PI) which leads to mature GPI membrane anchor precursors, which are transferred to certain proteins in the endoplasmic reticulum. The substrate specificities of the GlcNAc-PI de-N-acetylase activities of African trypanosomes and human (HeLa) cells were studied with respect to the N-acyl groups (R) that could be removed from a series of GlcNR-PI substrates, where R = acetyl (Ac), propionyl (Pr), butyryl (Bu), isobutyryl (iBu), pentanoyl (Pen) or hexanoyl (Hex). The data show that the trypanosomal and HeLa enzymes had similar specificities and that the turnover of GlcNR-PIs by the trypanosomal enzyme was in the order GlcNAc-PI > GlcNPr-PI GlcNBu - PI ≈ GlcNiBu - PI ≈ GlcNPen - PI
GlcNHex - PI. The trypanosome and HeLa de-N-acetylases were unable to de-N-acetylate mannosylated GlcNAc-PI intermediates, which explains why de-N-acetylation must precede mannosylation in the GPI biosynthetic pathway.
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November 1997
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Research Article|
November 15 1997
Substrate specificity of the N-acetylglucosaminyl-phosphatidylinositol de-N-acetylase of glycosylphosphatidylinositol membrane anchor biosynthesis in African trypanosomes and human cells Available to Purchase
K. Deepak SHARMA;
K. Deepak SHARMA
*Department of Biochemistry, Carbohydrate Research Centre, University of Dundee, Dundee DD1 4HN, Scotland, U.K.
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K. Terry SMITH;
K. Terry SMITH
*Department of Biochemistry, Carbohydrate Research Centre, University of Dundee, Dundee DD1 4HN, Scotland, U.K.
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Arthur CROSSMAN;
Arthur CROSSMAN
†Department of Chemistry, Carbohydrate Research Centre, University of Dundee, Dundee DD1 4HN, Scotland, U.K.
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S. John BRIMACOMBE;
S. John BRIMACOMBE
†Department of Chemistry, Carbohydrate Research Centre, University of Dundee, Dundee DD1 4HN, Scotland, U.K.
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A. J. Michael FERGUSON
A. J. Michael FERGUSON
1
*Department of Biochemistry, Carbohydrate Research Centre, University of Dundee, Dundee DD1 4HN, Scotland, U.K.
1To whom correspondence should be addressed.
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Publisher: Portland Press Ltd
Received:
June 12 1997
Accepted:
July 18 1997
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1997
1997
Biochem J (1997) 328 (1): 171–177.
Article history
Received:
June 12 1997
Accepted:
July 18 1997
Citation
K. Deepak SHARMA, K. Terry SMITH, Arthur CROSSMAN, S. John BRIMACOMBE, A. J. Michael FERGUSON; Substrate specificity of the N-acetylglucosaminyl-phosphatidylinositol de-N-acetylase of glycosylphosphatidylinositol membrane anchor biosynthesis in African trypanosomes and human cells. Biochem J 15 November 1997; 328 (1): 171–177. doi: https://doi.org/10.1042/bj3280171
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