Expression and characterization of recombinant kurtoxin, an inhibitor of T-type voltage-gated calcium channels
| Autor: | Young-Jae Eu, Hye Jung Min, Seung-Yeol Nah, Jun Ho Lee, Chul Won Lee, Jae Il Kim, Ha Hyung Kim, Eun-Mi Cho, Kenton J. Swartz |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Protein Folding
Protein Conformation Molecular Sequence Data Biophysics Scorpion Venoms Peptide Biochemistry Protein Structure Secondary law.invention Calcium Channels T-Type chemistry.chemical_compound law Escherichia coli Amino Acid Sequence Cysteine Molecular Biology Protein secondary structure chemistry.chemical_classification Scorpion toxin Voltage-dependent calcium channel Chemistry Calcium channel T-type calcium channel Cell Biology Recombinant Proteins Recombinant DNA Kurtoxin Oxidation-Reduction |
| Zdroj: | Biochemical and Biophysical Research Communications. 416:277-282 |
| ISSN: | 0006-291X |
| DOI: | 10.1016/j.bbrc.2011.11.003 |
| Popis: | Kurtoxin, a 63-amino acid peptide stabilized by four disulfide bonds, is the first reported peptide inhibitor of T-type voltage-gated calcium channels. Although T-type calcium channels have been implicated in a number of disease states, including epilepsy, chronic pain, hypertension and cancer, the lack of selective inhibitors has slowed progress in understanding their precise roles. Kurtoxin is a potentially valuable tool with which to study T-type calcium channels. However, because of the limited availability of the native protein, little is known about the structure and molecular mechanism of kurtoxin. Here we report the expression of kurtoxin in Escherichia coli and the structural and functional characterization of the recombinant protein. The disulfide bond pairings and secondary structure of recombinant kurtoxin were characterized through enzymatic cleavage, mass analysis and CD spectroscopy. Recombinant kurtoxin almost completely inhibited the T-type calcium channel in a manner identical to the native toxin. The availability of recombinant kurtoxin that is identical to the native toxin should help in the study of T-type calcium channels and enable development of new strategies for producing even more-selective T-type calcium channel inhibitors and for investigating the molecular basis of the toxin-channel interactions. |
| Databáze: | OpenAIRE |
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