The complete cDNA sequence and expression of the first major allergenic protein of Malassezia furfur, Mal f 1.
For the first time the complete cDNA encoding a major allergen and novel protein of the yeast Malassezia furfur, Mal f 1, has been sequenced and expressed. The amino acid sequences of nine tryptic peptides of the protein were determined. Oligonucleotides were designed from these amino acid sequences. The cDNA sequence was obtained by hybridizing these primers to mRNA and enhancement by reverse-transcriptase PCR techniques. The cDNA is 1176 bp in length. It shows an open reading frame of 1050 bp coding for a protein of 38178 Da and a deduced amino acid sequence containing 350 residues. The hydropathy plot and the tryptic digest indicate that the first 22 amino acids represent a leader sequence determining a mature protein of 35 988 Da. The complete encoding cDNA was expressed as a maltose-binding protein fusion protein in Escherichia coli. The recombinant fusion protein reacted with our specific monoclonal antibody and with IgE from patients with atopic dermatitis.
Schmidt
M
M
Department of Laboratory Medicine, Karolinska Hospital, Stockholm, Sweden.
Zargari
A
A
Holt
P
P
Lindbom
L
L
Hellman
U
U
Whitley
P
P
van der Ploeg
I
I
Härfast
B
B
Scheynius
A
A
D000485:Allergens / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary; D003876:Dermatitis, Atopic / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D006801:Humans; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000276:immunology; D008290:Malassezia / Q000737:chemistry*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D014357:Trypsin
D000485:Allergens; D000911:Antibodies, Monoclonal; D017931:DNA Primers; D018076:DNA, Complementary; D005656:Fungal Proteins; C106960:Mal f 1 allergen; D010446:Peptide Fragments; D007073:Immunoglobulin E; D014357:Trypsin