Characterization of the agent of "high plains disease": mass spectrometry determines the sequence of the disease-specific protein

The "32-kDa" protein specifically associated with high plains disease was characterized by time-of-flight mass spectrometry, after the agent had been isolated in pure culture by "vascular puncture inoculation," a novel mechanical means of transmission. Two isolates from different...

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Bibliographic Details
Published in:The Journal of biological chemistry 2004-01, Vol.279 (1), p.488-494
Main Authors: She, Yi-Min, Seifers, Dallas L, Haber, Steve, Ens, Werner, Standing, Kenneth G
Format: Article
Language:English
Subjects:
Altitude
Amino Acid Sequence
High Plains disease
High plains virus
Molecular Sequence Data
Peptide Fragments
Peptide Mapping
Plant Diseases
Plant Proteins - chemistry
Plant Viruses - chemistry
Plant Viruses - pathogenicity
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Triticum
United States
Zea mays
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Summary:The "32-kDa" protein specifically associated with high plains disease was characterized by time-of-flight mass spectrometry, after the agent had been isolated in pure culture by "vascular puncture inoculation," a novel mechanical means of transmission. Two isolates from different geographic locations each consisted of a mixture of subpopulations that were highly homologous to an amino acid sequence derived from a nucleotide sequence (U60141) deposited in GenBank trade mark by the Nebraska group as "the probable N-protein of high plains virus." However, the U60141 sequence was found to be incomplete; de novo sequencing of peptides produced by proteolytic digestions of the 32-kDa band from an SDS-PAGE separation showed that an additional 18 amino acid residues were present at the N terminus. BLAST (basic local alignment search tool) examination of the sequence showed no significant homology with any protein in the databases, indicating that the infectious agent of high plains disease is likely a member of a hitherto unclassified virus group.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M308506200