Abstract
Background
Malassezia globosa constitutes a part of the normal flora of human skin, but may induce IgE production
in atopic dermatitis (AD). However, information on M. globosa allergens is scant.
Objective
To identify the major M. globosa allergens by using proteomic analysis.
Methods
Immunoglobulin E (IgE) immunoblotting and cross-inhibition tests for M. globosa allergens were performed using sera from AD patients and control subjects. These
allergens were identified and characterized using the proteomics approach involving
a combination of two-dimensional (2D) electrophoresis, mass spectrometry, and bioinformatics
tools. We cloned the cDNA of this allergen using sequences obtained by 5′- and 3′-rapid
amplification of cDNA ends polymerase chain reaction.
Results
The sera of the AD patients had IgE-reactive 40–45-kDa protein components. By 2D immunoblotting,
we detected a 42-kDa protein spot with an isoelectric point (pI) of 4.8; the protein was highly reactive to IgE and was designated MGp42. Full-length
MGp42 cDNA contained a 1908-bp open reading frame encoding 635 amino acid residues
(calculated molecular mass, 69.7 kDa; pI, 6.02). The N-terminal MGp42 sequence started from the 250th residue (Asp-250) of
the deduced amino acid sequence and consisted of 386 amino acid residues; these results
are consistent with those of 2D immunoblotting. MGp42 showed sequence similarity to
members of the heat shock protein 70 (hsp70) family. Immunoblot inhibition tests revealed
no IgE cross-reactivity between MGp42 and human HSP70.
Conclusions
MGp42 may be a cleavage product of intact HSP70. This novel M. globosa allergen could be useful for the diagnosis of AD.
Keywords
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Article info
Publication history
Published online: July 12, 2010
Accepted:
May 20,
2009
Received in revised form:
April 11,
2009
Received:
December 4,
2008
Identification
Copyright
© 2009 Japanese Society for Investigative Dermatology. Published by Elsevier Inc. All rights reserved.
