Document Detail

Human xylosyltransferase I and N-terminal truncated forms: functional characterization of the core enzyme.
MedLine Citation:
PMID:  16225459     Owner:  NLM     Status:  MEDLINE    
Human XT-I (xylosyltransferase I; EC initiates the biosynthesis of the glycosaminoglycan linkage region and is a diagnostic marker of an enhanced proteoglycan biosynthesis. In the present study, we have investigated mutant enzymes of human XT-I and assessed the impact of the N-terminal region on the enzymatic activity. Soluble mutant enzymes of human XT-I with deletions at the N-terminal domain were expressed in insect cells and analysed for catalytic activity. As many as 260 amino acids could be truncated at the N-terminal region of the enzyme without affecting its catalytic activity. However, truncation of 266, 272 and 273 amino acids resulted in a 70, 90 and >98% loss in catalytic activity. Interestingly, deletion of the single 12 amino acid motif G261KEAISALSRAK272 leads to a loss-of-function XT-I mutant. This is in agreement with our findings analysing the importance of the Cys residues where we have shown that C276A mutation resulted in a nearly inactive XT-I enzyme. Moreover, we investigated the location of the heparin-binding site of human XT-I using the truncated mutants. Heparin binding was observed to be slightly altered in mutants lacking 289 or 568 amino acids, but deletion of the potential heparin-binding motif P721KKVFKI727 did not lead to a loss of heparin binding capacity. The effect of heparin or UDP on the XT-I activity of all mutants was not significantly different from that of the wild-type. Our study demonstrates that over 80% of the nucleotide sequence of the XT-I-cDNA is necessary for expressing a recombinant enzyme with full catalytic activity.
Sandra Müller; Jennifer Disse; Manuela Schöttler; Sylvia Schön; Christian Prante; Thomas Brinkmann; Joachim Kuhn; Knut Kleesiek; Christian Götting
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Biochemical journal     Volume:  394     ISSN:  1470-8728     ISO Abbreviation:  Biochem. J.     Publication Date:  2006 Feb 
Date Detail:
Created Date:  2006-01-26     Completed Date:  2006-03-24     Revised Date:  2013-06-07    
Medline Journal Info:
Nlm Unique ID:  2984726R     Medline TA:  Biochem J     Country:  England    
Other Details:
Languages:  eng     Pagination:  163-71     Citation Subset:  IM    
Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum Nordrhein-Westfalen, Universitätsklinik der Ruhr-Universität Bochum, Georgstrasse 11, 32545 Bad Oeynhausen, Germany.
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MeSH Terms
Amino Acid Sequence
Cell Line
Molecular Sequence Data
Mutagenesis, Site-Directed
Pentosyltransferases / chemistry*,  genetics,  metabolism*
Protein Binding
Sequence Alignment
Sequence Deletion / genetics*
Sequence Homology, Amino Acid
Reg. No./Substance:
9005-49-6/Heparin; EC 2.4.2.-/Pentosyltransferases; EC xylose-protein xylosyltransferase

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