Isolation and characterization of the early nodule-specific protein homologue (Hev b 13), an allergenic lipolytic esterase from Hevea brasiliensis latex.
Recurring reports of a highly allergenic 42-46-kDa protein in Hevea brasiliensis latex appeared to have been resolved with the discovery of a 43-kDa allergenic latex protein that was a homologue to patatin. However, the low to moderate prevalence of sensitization to the protein, designated Hev b 7, among latex-allergic patients could not adequately explain the frequent observations of the 42-46-kDa allergen. This led to the hypothesis that another, more allergenic protein of a similar molecular mass existed in Hevea latex. We report the isolation and purification of a 42.98-kDa latex glycoprotein showing homology to the early nodule-specific protein (ENSP) of the legumes Medicago sativa, Medicago truncatula, and Glycine max. The protein is allergenic, being recognized by immunoglobulin E (IgE) in sera from latex-allergic patients. The IgE epitope resides on the carbohydrate moiety of the protein, and the presence of a similar carbohydrate component on potato tuber patatin enables the latter to inhibit IgE binding to the ENSP homologue. The cDNA encoding the ENSP homologue was isolated by reverse transcription-PCR and cloned. The protein predicted from the cDNA sequence has 391 amino acids, the first 26 of which constitute a putative signal peptide. The deduced molecular mass of the mature protein is 40.40 kDa, while its isoelectric point is estimated at 5.0. The discrepancy between the predicted and observed molecular mass might be due to glycosylation, for which three N-sites on the protein are predicted. The purified protein showed lipase and esterase activities and may be involved in plant defense.
Arif
Siti Arija M
SA
Rubber Research Institute of Malaysia, 50450 Kuala Lumpur, Malaysia. sitiarija@lgm.gov.my
Hamilton
Robert G
RG
Yusof
Faridah
F
Chew
Nyu-Ping
NP
Loke
Yin-Ho
YH
Nimkar
Subodh
S
Beintema
Jaap J
JJ
Yeang
Hoong-Yeet
HY
D000485:Allergens / Q000737:chemistry* / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D001483:Base Sequence; D015153:Blotting, Western; D002241:Carbohydrates / Q000737:chemistry; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D004950:Esterases / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007073:Immunoglobulin E / Q000737:chemistry; D007525:Isoelectric Focusing; D007840:Latex / Q000737:chemistry* / Q000378:metabolism; D008049:Lipase / Q000378:metabolism; D029902:Medicago / Q000378:metabolism; D000455:Medicago sativa / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D010940:Plant Proteins; D011485:Protein Binding; D021382:Protein Sorting Signals; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013025:Glycine max / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D014357:Trypsin / Q000494:pharmacology
D000485:Allergens; D052179:Antigens, Plant; D002241:Carbohydrates; D018076:DNA, Complementary; D000939:Epitopes; C473106:Hev b 13 allergen, Hevea brasiliensis; D007840:Latex; D010455:Peptides; D010940:Plant Proteins; D021382:Protein Sorting Signals; D011506:Proteins; D012333:RNA, Messenger; D007073:Immunoglobulin E; D004247:DNA; D004950:Esterases; D008049:Lipase; D014357:Trypsin