Nucleotide sequence analysis of three cDNAs coding for Poa p IX isoallergens of Kentucky bluegrass pollen.
Grass pollen allergens are one of the major causes of type I allergic reactions (allergic rhinoconjunctivitis, allergic bronchial asthma, and hayfever) in temperate climates afflicting 15-20% of a genetically predisposed population. Workers have found considerable physico- and immunochemical heterogeneity within the grass pollen allergens which has made them difficult to purify for both therapeutic uses and further biochemical study. We recently reported the construction of a cDNA library in lambda gt11 using mRNA extracted from dehydrated Kentucky bluegrass (KBG, Poa pratensis). Here, we present the nucleotide and deduced amino acid sequences for three KBG pollen allergen cDNA clones, KBG 41, 60, and 31, which were isolated from the above library using a pool of six sera from grass pollen allergic patients. These clones exhibit an exceptionally high degree of sequence similarity to one another, only minor similarity to other known allergens, and no homologies to other known proteins or genes. The predicted molecular mass for the cloned proteins range from 28.3 to 37.8 kDa with pI values of 9.6-10.2. All three clones appear to possess leader peptides and lack asparagine sequons required for N-glycosylation. Therefore, the molecular mass of the post-translationally modified proteins were calculated to be 28.4-34.9 kDa, which is consistent with the size of the polypeptides revealed in Western blots of pollen proteins using an antiserum to a recombinant peptide encoded by the partial cDNA clone KBG 8.3. Northern blotting analysis indicates that expression of the genes corresponding to these clones is confined to pollen tissue. The results suggest that the clones code for a group of proteins that represent a new and previously uncharacterized group of grass pollen isoallergens, which have been hereby designated as Poa p IX.
Silvanovich
A
A
Department of Plant Science, University of Manitoba, Winnipeg, Canada.
Astwood
J
J
Zhang
L
L
Olsen
E
E
Kisil
F
F
Sehon
A
A
Mohapatra
S
S
Hill
R
R
D000485:Allergens / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002999:Clone Cells; D004247:DNA / Q000235:genetics*; D015870:Gene Expression; D014644:Genetic Variation; D006801:Humans; D008969:Molecular Sequence Data; D006109:Poaceae; D011058:Pollen; D012313:RNA / Q000235:genetics; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D000485:Allergens; D012313:RNA; D004247:DNA