Publication:
Mass spectrometric analysis of electrophoretically separated allergens and proteases in grass pollen diffusates

dc.contributor.author Raftery, Mark J en_US
dc.contributor.author Saldanha, Rohit Gregory en_US
dc.contributor.author Geczy, Carolyn L en_US
dc.contributor.author Kumar, Rakesh K en_US
dc.date.accessioned 2021-11-25T12:38:18Z
dc.date.available 2021-11-25T12:38:18Z
dc.date.issued 2003 en_US
dc.description.abstract Background: Pollens are important triggers for allergic asthma and seasonal rhinitis, and proteases released by major allergenic pollens can injure airway epithelial cells in vitro. Disruption of mucosal epithelial integrity by proteases released by inhaled pollens could promote allergic sensitisation. Methods: Pollen diffusates from Kentucky blue grass (Poa pratensis), rye grass (Lolium perenne) and Bermuda grass (Cynodon dactylon) were assessed for peptidase activity using a fluorogenic substrate, as well as by gelatin zymography. Following one- or two-dimensional gel electrophoresis, Coomassie-stained individual bands/spots were excised, subjected to tryptic digestion and analysed by mass spectrometry, either MALDI reflectron TOF or microcapillary liquid chromatography MS-MS. Database searches were used to identify allergens and other plant proteins in pollen diffusates. Results: All pollen diffusates tested exhibited peptidase activity. Gelatin zymography revealed high Mr proteolytic activity at ~95,000 in all diffusates and additional proteolytic bands in rye and Bermuda grass diffusates, which appeared to be serine proteases on the basis of inhibition studies. A proteolytic band at Mr ~35,000 in Bermuda grass diffusate, which corresponded to an intense band detected by Western blotting using a monoclonal antibody to the timothy grass (Phleum pratense) group 1 allergen Phl p 1, was identified by mass spectrometric analysis as the group 1 allergen Cyn d 1. Two-dimensional analysis similarly demonstrated proteolytic activity corresponding to protein spots identified as Cyn d 1. Conclusion: One- and two-dimensional electrophoretic separation, combined with analysis by mass spectrometry, is useful for rapid determination of the identities of pollen proteins. A component of the proteolytic activity in Bermuda grass diffusate is likely to be related to the allergen Cyn d 1. en_US
dc.identifier.issn 1465-9921 en_US
dc.identifier.uri http://hdl.handle.net/1959.4/10186
dc.language English
dc.language.iso EN en_US
dc.rights CC BY-NC-ND 3.0 en_US
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/3.0/au/ en_US
dc.source Legacy MARC en_US
dc.subject.other mass spectrometry en_US
dc.subject.other pollens en_US
dc.subject.other proteomics en_US
dc.subject.other Respiratory Diseases (321027) en_US
dc.subject.other Pathology (321020) en_US
dc.subject.other Allergy (320201) en_US
dc.title Mass spectrometric analysis of electrophoretically separated allergens and proteases in grass pollen diffusates en_US
dc.type Journal Article en
dcterms.accessRights metadata only access
dspace.entity.type Publication en_US
unsw.accessRights.uri http://purl.org/coar/access_right/c_14cb
unsw.identifier.doiPublisher http://dx.doi.org/doi:10.1186/1465-9921-4-10 en_US
unsw.relation.faculty Medicine & Health
unsw.relation.ispartofissue 10 en_US
unsw.relation.ispartofjournal Respiratory Research en_US
unsw.relation.ispartofpagefrompageto 1-12 en_US
unsw.relation.ispartofvolume 4 en_US
unsw.relation.originalPublicationAffiliation Raftery, Mark J, Medical Sciences, Faculty of Medicine, UNSW en_US
unsw.relation.originalPublicationAffiliation Saldanha, Rohit Gregory, Medical Sciences, Faculty of Medicine, UNSW en_US
unsw.relation.originalPublicationAffiliation Geczy, Carolyn L, Medical Sciences, Faculty of Medicine, UNSW en_US
unsw.relation.originalPublicationAffiliation Kumar, Rakesh K, Medical Sciences, Faculty of Medicine, UNSW en_US
unsw.relation.school School of Medical Sciences *
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