Linked Life Data

12182702

Source:http://linkedlifedata.com/resource/pubmed/id/12182702

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
4
pubmed:dateCreated
2002-8-16
pubmed:abstractText
Functional analysis of the hyperglycosylated arabinogalactan-proteins (AGPs) attempts to relate biological roles to the molecular properties that result largely from O-Hyp glycosylation putatively coded by the primary sequence. The Hyp contiguity hypothesis predicts contiguous Hyp residues as attachment sites for arabino-oligosaccharides (arabinosides) and clustered, non-contiguous Hyp residues as arabinogalactan polysaccharide sites. Although earlier tests of naturally occurring hydroxyproline-rich glycoproteins (HRGPs) and HRGPs designed by synthetic genes were consistent with a sequence-driven code, the predictive value of the hypothesis starting from the DNA sequences of known AGPs remained untested due to difficulties in purifying a single AGP for analysis. However, expression in tobacco (Nicotiana tabacum) of the major tomato (Lycopersicon esculentum) AGP, LeAGP-1, as an enhanced green fluorescent protein fusion glycoprotein (EGFP)-LeAGP-1, increased its hydrophobicity sufficiently for chromatographic purification from other closely related endogenous AGPs. We also designed and purified two variants of LeAGP-1 for future functional analysis: one lacking the putative glycosylphosphatidylinositol (GPI)-anchor signal sequence; the other lacking a 12-residue internal lysine-rich region. Fluorescence microscopy of plasmolysed cells confirmed the location of LeAGP-1 at the plasma membrane outer surface and in Hechtian threads. Hyp glycoside profiles of the fusion glycoproteins gave ratios of Hyp-polysaccharides to Hyp-arabinosides plus non-glycosylated Hyp consistent with those predicted from DNA sequences by the Hyp contiguity hypothesis. These results demonstrate a route to the purification of AGPs and the use of the Hyp contiguity hypothesis for predicting the Hyp O-glycosylation profile of an HRGP from its DNA sequence.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0960-7412
pubmed:author
pubmed:issnType
Print
pubmed:volume
31
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
431-44
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:12182702-Amino Acid Sequence, pubmed-meshheading:12182702-Amino Acids, pubmed-meshheading:12182702-Base Sequence, pubmed-meshheading:12182702-Carbohydrate Metabolism, pubmed-meshheading:12182702-Cells, Cultured, pubmed-meshheading:12182702-Circular Dichroism, pubmed-meshheading:12182702-Gene Expression Regulation, Plant, pubmed-meshheading:12182702-Glycosylation, pubmed-meshheading:12182702-Glycosylphosphatidylinositols, pubmed-meshheading:12182702-Green Fluorescent Proteins, pubmed-meshheading:12182702-Hydroxyproline, pubmed-meshheading:12182702-Luminescent Proteins, pubmed-meshheading:12182702-Lycopersicon esculentum, pubmed-meshheading:12182702-Membrane Glycoproteins, pubmed-meshheading:12182702-Models, Molecular, pubmed-meshheading:12182702-Molecular Sequence Data, pubmed-meshheading:12182702-Mucoproteins, pubmed-meshheading:12182702-Plant Proteins, pubmed-meshheading:12182702-Plants, Genetically Modified, pubmed-meshheading:12182702-Protein Conformation, pubmed-meshheading:12182702-Recombinant Fusion Proteins, pubmed-meshheading:12182702-Tobacco
pubmed:year
2002
pubmed:articleTitle
Tomato LeAGP-1 arabinogalactan-protein purified from transgenic tobacco corroborates the Hyp contiguity hypothesis.
pubmed:affiliation
Department of Chemistry and Biochemistry, Ohio University, Athens, OH 45710, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, Non-P.H.S., Research Support, Non-U.S. Gov't