Document Detail

Structure of a hydroxyproline (Hyp)-arabinogalactan polysaccharide from repetitive Ala-Hyp expressed in transgenic Nicotiana tabacum.
MedLine Citation:
PMID:  14724279     Owner:  NLM     Status:  MEDLINE    
A synthetic gene encoding the fusion protein (Ala-Hyp)(51)-enhanced green fluorescent protein expressed in Nicotiana tabacum cells produced a fusion glycoprotein with all proline residues hydroxylated and substituted with an arabinogalactan polysaccharide. Alkaline hydrolysis of the fusion glycoprotein yielded a population of hydroxyproline (Hyp)-arabinogalactan polysaccharides ranging in size from 13 to 26 saccharide residues/Hyp, with a median size of 15-17 residues. We isolated a 15-residue Hyp-arabinogalactan for structure determination by sugar analyses and one- and two-dimensional nuclear magnetic resonance techniques that provided the assignment of proton and carbon signals of a small polysaccharide O-linked to the hydroxyl group of Hyp. The polysaccharide consisted of a 1,3-linked beta-D-Galp backbone with a single 1,6-linked beta-D-Galp "kink." The backbone had two side chains of Galp substituted at position 3 with an arabinose di- or trisaccharide and at position 6 with glucuronic acid or rhamnosyl glucuronic acid. Energy-minimized space-filling molecular models showed hydrogen bonding within polysaccharides attached to repetitive Ala-Hyp and also between polysaccharides and the peptide backbone. Polysaccharides distorted the peptide Ramachandran angles consistent with the circular dichroic spectra of isolated (Ala-Hyp)(51) and its reversion to a polyproline II-like helix after deglycosylation. This first complete structure of a Hyp-arabinogalactan polysaccharide shows that computer-based molecular modeling of Hyp-rich glycoproteins is now feasible and supports the suggestion that small repetitive subunits comprise larger arabinogalactan polysaccharides.
Li Tan; Feng Qiu; Derek T A Lamport; Marcia J Kieliszewski
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.     Date:  2004-01-14
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  279     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2004 Mar 
Date Detail:
Created Date:  2004-03-22     Completed Date:  2004-05-07     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  13156-65     Citation Subset:  IM    
Department of Chemistry and Biochemistry, Ohio University, Athens, OH 45701, USA.
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MeSH Terms
Carbon / chemistry
Circular Dichroism
Galactans / chemistry*
Glucuronic Acid / chemistry
Green Fluorescent Proteins
Hydroxyproline / chemistry*
Luminescent Proteins / metabolism
Magnetic Resonance Spectroscopy
Models, Chemical
Models, Molecular
Polysaccharides / chemistry*
Proline / chemistry
Recombinant Fusion Proteins / chemistry
Tobacco / metabolism*
Grant Support
2P41 RR 05351-06/RR/NCRR NIH HHS
Reg. No./Substance:
0/Galactans; 0/Luminescent Proteins; 0/Polysaccharides; 0/Recombinant Fusion Proteins; 147-85-3/Proline; 147336-22-9/Green Fluorescent Proteins; 51-35-4/Hydroxyproline; 576-37-4/Glucuronic Acid; 7440-44-0/Carbon; 9036-66-2/arabinogalactan

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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