Characterisation of the rubber elongation factor from ammoniated latex by electrophoresis and mass spectrometry.
Rubber elongation factor (REF) is considered as one of the major allergens present in latex. An extraction and purification protocol for preparation of REF standards has been modified. A protein fraction was extracted from ammoniated latex sap and purified by gel filtration chromatography. The purified and concentrated proteins were separated by sodium dodecylsulfate-polyacrylamide gel electrophoresis into two major bands. These bands were further characterised by matrix-assisted laser desorption/ionisation time-of-flight and nano-electrospray ionization mass spectrometry. REF and a truncated form could be ascertained by the mass and fragmentation pattern of the tryptic peptides. In the faster migrating band an additional peptide could be identified. This peptide is also present in Hevb3 and a Mr 27000 latex allergen. Our findings indicate that conventional REF preparations as standards may contain additional allergenic proteins.
Dürauer
A
A
Institute of Applied Microbiology, University of Agricultural Sciences, Vienna, Austria.
Csaszar
E
E
Mechtler
K
K
Jungbauer
A
A
Schmid
E
E
D000485:Allergens; D000941:Antigens / Q000032:analysis / Q000302:isolation & purification; D052179:Antigens, Plant; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D007840:Latex / Q000737:chemistry*; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D010940:Plant Proteins / Q000032:analysis / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D000485:Allergens; D000941:Antigens; D052179:Antigens, Plant; D007840:Latex; D010446:Peptide Fragments; D010940:Plant Proteins; C061477:REF protein, Hevea brasiliensis