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 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.
She
Yi-Min
YM
Department of Physics & Astronomy, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada.
Seifers
Dallas L
DL
Haber
Steve
S
Ens
Werner
W
Standing
Kenneth G
KG
D000531:Altitude; D000595:Amino Acid Sequence; D008969:Molecular Sequence Data; D010446:Peptide Fragments; D010449:Peptide Mapping; D010935:Plant Diseases; D010940:Plant Proteins / Q000737:chemistry*; D010942:Plant Viruses / Q000737:chemistry / Q000472:pathogenicity*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014908:Triticum; D014481:United States; D003313:Zea mays
D010446:Peptide Fragments; D010940:Plant Proteins