Purification and characterization of Alt a-29 from Alternaria alternata.
A major protein component reactive with pooled human atopic sera was isolated from a lyophilized broth extract of Alternaria alternata 34-016. By successive chromatography on Whatman DE-52, Sephadex G-100 and Mono Q HR5/5, a low molecular weight antigen was obtained. Comparison with standard proteins on Sephadex G-100 indicated its molecular weight was 31 kD. Non-reduced samples run on SDS-PAGE showed a band at 29.2 kD which reacted strongly with human IgE. After reduction, it produced a doublet pattern on SDS-PAGE with MW 14.5 and 16.0 kD. The doublet pattern was confirmed by Western blotting with pooled human atopic sera. IEF of the protein showed a major component with a PI of 4.15 and two minor components at 4.25 and 4.40. Immunoblots of the IEF bands showed all three were reactive with human IgE. Ion exchange chromatography of the protein on Mono Q HR5/5 resulted in three resolved components, all of which are immunoreactive. Together with the IEF data, this suggests that there are several conformational or structural isoforms of this protein.
Curran
I H
IH
Bureau of Drug Research, Health Protection Branch, Health and Welfare, Ottawa, Canada.
Young
N M
NM
Burton
M
M
Vijay
H M
HM
D000485:Allergens / Q000276:immunology / Q000302:isolation & purification*; D000528:Alternaria / Q000737:chemistry / Q000276:immunology*; D000595:Amino Acid Sequence; D005656:Fungal Proteins / Q000737:chemistry* / Q000276:immunology*; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology; D007073:Immunoglobulin E / Q000276:immunology; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight
D000485:Allergens; D005656:Fungal Proteins; D007073:Immunoglobulin E