Expression and Localization of an Hsp70 Protein in the Microsporidian Encephalitozoon cuniculi
Microsporidia spore surface proteins are an important, under investigated aspect of spore/host cell attachment and infection. For comparison analysis of surface proteins, we required an antibody control specific for an intracellular protein. An endoplasmic reticulum-associated heat shock protein 7...
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| Format: | Article |
| Language: | English |
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Wiley
2010-01-01
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| Series: | International Journal of Microbiology |
| Online Access: | http://dx.doi.org/10.1155/2010/523654 |
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| author | Carrie E. Jolly Cory A. Leonard J. Russell Hayman |
| author_facet | Carrie E. Jolly Cory A. Leonard J. Russell Hayman |
| author_sort | Carrie E. Jolly |
| collection | DOAJ |
| description | Microsporidia spore surface proteins are an important, under investigated aspect of spore/host cell attachment and infection. For comparison analysis of surface proteins, we required an antibody control specific for an intracellular protein. An endoplasmic reticulum-associated heat shock protein 70 family member (Hsp70; ECU02_0100; “C1”) was chosen for further analysis. DNA encoding the C1 hsp70 was amplified, cloned and used to heterologously express the C1 Hsp70 protein, and specific antiserum was generated. Two-dimensional Western blotting analysis showed that the purified antibodies were monospecific. Immunoelectron microscopy of developing and mature E. cuniculi spores revealed that the protein localized to internal structures and not to the spore surface. In spore adherence inhibition assays, the anti-C1 antibodies did not inhibit spore adherence to host cell surfaces, whereas antibodies to a known surface adhesin (EnP1) did so. In future studies, the antibodies to the ‘C1’ Hsp70 will be used to delineate spore surface protein expression. |
| format | Article |
| id | doaj-art-ea5aabf757124d128cb42c4a30123507 |
| institution | Kabale University |
| issn | 1687-918X 1687-9198 |
| language | English |
| publishDate | 2010-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | International Journal of Microbiology |
| spelling | doaj-art-ea5aabf757124d128cb42c4a301235072025-02-03T06:46:25ZengWileyInternational Journal of Microbiology1687-918X1687-91982010-01-01201010.1155/2010/523654523654Expression and Localization of an Hsp70 Protein in the Microsporidian Encephalitozoon cuniculiCarrie E. Jolly0Cory A. Leonard1J. Russell Hayman2Department of Microbiology, James H. Quillen College of Medicine, East Tennessee State University, Box 70579, Johnson City, TN 37614, USADepartment of Microbiology, James H. Quillen College of Medicine, East Tennessee State University, Box 70579, Johnson City, TN 37614, USADepartment of Microbiology, James H. Quillen College of Medicine, East Tennessee State University, Box 70579, Johnson City, TN 37614, USAMicrosporidia spore surface proteins are an important, under investigated aspect of spore/host cell attachment and infection. For comparison analysis of surface proteins, we required an antibody control specific for an intracellular protein. An endoplasmic reticulum-associated heat shock protein 70 family member (Hsp70; ECU02_0100; “C1”) was chosen for further analysis. DNA encoding the C1 hsp70 was amplified, cloned and used to heterologously express the C1 Hsp70 protein, and specific antiserum was generated. Two-dimensional Western blotting analysis showed that the purified antibodies were monospecific. Immunoelectron microscopy of developing and mature E. cuniculi spores revealed that the protein localized to internal structures and not to the spore surface. In spore adherence inhibition assays, the anti-C1 antibodies did not inhibit spore adherence to host cell surfaces, whereas antibodies to a known surface adhesin (EnP1) did so. In future studies, the antibodies to the ‘C1’ Hsp70 will be used to delineate spore surface protein expression.http://dx.doi.org/10.1155/2010/523654 |
| spellingShingle | Carrie E. Jolly Cory A. Leonard J. Russell Hayman Expression and Localization of an Hsp70 Protein in the Microsporidian Encephalitozoon cuniculi International Journal of Microbiology |
| title | Expression and Localization of an Hsp70 Protein in the Microsporidian Encephalitozoon cuniculi |
| title_full | Expression and Localization of an Hsp70 Protein in the Microsporidian Encephalitozoon cuniculi |
| title_fullStr | Expression and Localization of an Hsp70 Protein in the Microsporidian Encephalitozoon cuniculi |
| title_full_unstemmed | Expression and Localization of an Hsp70 Protein in the Microsporidian Encephalitozoon cuniculi |
| title_short | Expression and Localization of an Hsp70 Protein in the Microsporidian Encephalitozoon cuniculi |
| title_sort | expression and localization of an hsp70 protein in the microsporidian encephalitozoon cuniculi |
| url | http://dx.doi.org/10.1155/2010/523654 |
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