Cloning and characterization of the cDNA encoding apis mellifera (honeybee) venom protease / by Karen M. Winningham.
| Author/creator | Winningham, Karen M. author. |
| Other author | Hoffman, Donald R., degree supervisor. |
| Other author | Schmidt, Margit, degree supervisor. |
| Other author | East Carolina University. Department of Biology. |
| Format | Theses and dissertations |
| Production | 2001. |
| Description | 57 leaves : illustrations (some color) ; 28 cm |
| Supplemental Content | Access via ScholarShip |
| Subjects |
| Summary | Allergic reactions to Hymenoptera stinging insects have been reported in approximately 0.4 % (> 14 million) of the United States population. The allergic reactions to Hymenoptera venom range from fairly mild to extremely severe resulting in nearly 40 fatalities per year. Apis mellifera or honeybees are a major cause of stinging insect venom allergy. The composition of honeybee venom has been studied in great detail for almost 50 years; however, the venom proteins have not all been characterized. The most important allergens found in honeybee venom are the enzymes phospholipase A2 and hyaluronidase. Well-characterized secondary allergens include acid phosphatase and tetramers of the peptide melittin. A 39 kD protease was isolated from honeybee venom using gel filtration, cation exchange and reversed phase chromatography. Tryptic and Staphylococcal V8 protease digestions were used to determine the N-terminal sequence as well as a number of internal peptide sequences. The venom proteases isolated from bumblebee and paper wasp have proven to be important allergens, but the allergenic importance of the honeybee venom protease has not been assessed. On the basis of the partial peptide sequences obtained, experiments using reverse transcriptase mediated polymerase chain reactions (RT-PCR) and rapid amplification of cDNA ends (RACE) led to the full-length cDNA sequence. Analysis of the deduced amino acid sequence of the honeybee venom protease revealed the presence of an unexpected CUB domain. CUB domains are thought to be important in developmental processes and protein-protein interactions. Amino acid sequence analysis also showed that the honeybee venom protease contained an unusual methionine residue at the entrance of the substrate-binding pocket. The structural descriptions obtained from the cDNA sequence will aid in future experiments used to determine the allergenicity of the honeybee venom protease. |
| General note | Presented to the faculty of the Department of Biology. |
| General note | Advisor: Donald R. Hoffman |
| General note | Advisor: Margit Schmidt |
| Dissertation note | M.S. East Carolina University 2001 |
| Bibliography note | Includes bibliographical references (leaves 55-57). |
| Genre/form | dissertations. |
| Genre/form | Academic theses. |
| Genre/form | Academic theses. |
| Genre/form | Thèses et écrits académiques. |
Availability
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|---|---|---|---|---|
| Joyner | University Archives | ASK AT SPECIAL COLLECTIONS DESK | ✔ Available | Request Material |
| Electronic Resources | Access Content Online | ✔ Available |