Cloning, expression and characterization of two new IgE‐binding proteins from the yeast Malassezia sympodialis with sequence similarities to heat shock proteins and manganese superoxide dismutase

FEBS Journal - Tập 271 Số 10 - Trang 1885-1894 - 2004
Ann‐Catrin Andersson1, Omid Rasool1, Margit Schmidt2, Rimantas Kodzius3, Sabine Flückiger3, Arezou Zargari1, Reto Crameri3, Annika Scheynius1
1Department of Medicine, Clinical Allergy Research Unit, Karolinska Institutet and Hospital, Stockholm, Sweden
2Department of Biology and Department of Pathology and Laboratory Medicine, East Carolina University, Greenville, NC, USA
3Swiss Institute of Allergy and Asthma Research (SIAF), Davos, Switzerland

Tóm tắt

Malassezia sympodialis is an opportunistic yeast that colonizes human skin and may induce IgE and T cell reactivity in patients with atopic eczema/dermatitis syndrome (AEDS). Previously, we have cloned and expressed six recombinant allergens (rMala s 1 and rMala s 5 to rMala s 9) from this yeast. By combining high throughput screening and phage surface display techniques, 27 complete and partial IgE‐binding clones of M. sympodialis have been identified. Here we enlarged the panel of recombinant M. sympodialis allergens by RACE–PCR, cloning and nucleotide sequencing to obtain the coding sequences of two new IgE‐binding clones. The coding sequences of one of the clones showed similarity to the heat shock protein (HSP) family and the other to manganese superoxide dismutase (MnSOD), and both had a high degree of homology to human counterparts. The coding sequences were expressed in Escherichia coli as six‐histidine tagged recombinant proteins and generated products with molecular masses of 86.1 kDa for HSP and 22.4 kDa for MnSOD. Their IgE‐binding frequencies were shown to be 69% and 75%, respectively, to 28 sera from AEDS patients with serum IgE to M. sympodialis extract, indicating that HSP and MnSOD are major M. sympodialis allergens. In inhibition immunoblotting, M. sympodialis extract could inhibit the binding of serum IgE from AEDS patients to rHSP and rMnSOD in a concentration‐dependent manner. The high frequency of sera from AEDS patients, showing IgE binding to both HSP and MnSOD, indicates that these allergens, designated Mala s 10 and Mala s 11, could play a role in AEDS.

Từ khóa


Tài liệu tham khảo

10.1007/BF00399623

10.1128/JCM.38.5.1869-1875.2000

10.1128/JCM.40.4.1363-1367.2002

10.1128/JCM.41.10.4695-4699.2003

10.1034/j.1398-9995.1999.00089.x

10.1128/CMR.8.4.462

10.1128/CMR.5.2.101

10.1159/000053860

10.1046/j.1523-1747.2002.01758.x

10.1016/0190-9622(90)70100-V

10.1016/S1081-1206(10)62821-9

10.1111/j.1651-2227.1990.tb11467.x

10.1001/archderm.1990.01670290071011

10.1046/j.1365-2222.2000.00702.x

10.1046/j.1365-3083.1998.00271.x

10.1111/j.1365-2222.1995.tb03048.x

10.1080/000155500459303

10.1046/j.1523-1747.1999.00661.x

10.1111/j.1432-1033.1997.00181.x

10.1046/j.1432-1327.2000.01475.x

10.1046/j.1432-1327.1999.00247.x

10.1006/bbrc.1998.8944

10.1111/j.1348-0421.2000.tb02508.x

10.1046/j.1365-3164.2002.00291.x

10.1046/j.1365-2249.2002.01807.x

10.1159/000053664

10.1111/j.1365-2222.1997.tb00749.x

Hoffman C., 1997, Current Protocols in Molecular Biology, 13.11.12

10.1016/0378-1119(93)90253-Y

10.2174/1386207033329751

10.1093/nar/25.24.4876

Nicholas K.B., 1997, genedoc: Analysis and visualization of genetic variation, EMBNEW.NEWS., 4, 14

10.1084/jem.184.1.265

10.1111/j.1365-2133.1994.tb08532.x

10.1159/000233963

10.1016/0091-6749(94)90227-5

10.1111/j.1365-2222.1994.tb00985.x

10.1159/000063862

10.1016/S0140-6736(03)12193-9

10.1111/j.1398-9995.1998.tb03968.x

10.1159/000245983

10.1111/1523-1747.ep12343596

10.1111/j.1365-2222.1993.tb00310.x

Ghoreishi M., 2000, Heat shock proteins in the pathogenesis of inflammatory skin diseases, J. Med. Dent. Sci., 47, 143

10.1159/000237550

10.4049/jimmunol.168.3.1267

10.3109/10409238709083738

10.1159/000247298

Thông tin
Thông tin xuất bản

FEBS Journal

Tập 271 Số 10

1885-1894

Thông tin tác giả