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@proceedings{2281540, author = {Ravaszová, Karin and Hofrová, Alena and Hritz, Jozef}, booktitle = {XXII. Setkání biochemiků a molekulárních biologů}, keywords = {carbonic anhydrase IX, catalytic domain, protein tag}, language = {eng}, title = {Preparation of catalytic domain of carbonic anhydrase IX with different protein tags}, year = {2022} }
TY - CONF ID - 2281540 AU - Ravaszová, Karin - Hofrová, Alena - Hritz, Jozef PY - 2022 TI - Preparation of catalytic domain of carbonic anhydrase IX with different protein tags KW - carbonic anhydrase IX, catalytic domain, protein tag N2 - Carbonic anhydrase IX (CA IX) is a homodimeric transmembrane metalloenzyme, member of the α-carbonic anhydrase family. These enzymes catalyse (among other reactions) the interconversion between carbon dioxide and water to form bicarbonate and a proton and thus play an important role in pH maintenance. CA IX is overexpressed in many tumors helping them to maintain intracellular pH near physiological values while keeping the extracellular environment acidic and thus supports the cancer cells survival, migration and metastasis [1]. CA IX consists of 5 domains with catalytic and proteoglycan-like domain exposed in the extracellular part. The structural and biophysical properties of catalytic domain were determined in previous studies using mutant variants expressed in insect cells or yeast [2, 3]. We determined structural and biophysical properties of wild-type catalytic domain prepared with different expression and purification tags (thioredoxin, ubiquitin, NusA, ...) in E. coli cells. Proper characterization of extracellular part of CA IX is essential for design of selective inhibitors as α-carbonic anhydrase family reveals high degree of structural and functional similarity [4]. [1] S. Pastorekova and R. J. Gillies, “The role of carbonic anhydrase IX in cancer development: links to hypoxia, acidosis, and beyond,” Cancer and Metastasis Reviews, vol. 38, no. 1–2. Springer New York LLC, pp. 65–77, Jun. 15, 2019, doi: 10.1007/s10555-019-09799-0. [2] Alterio, Vincenzo et al. “Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX.” Proceedings of the National Academy of Sciences of the United States of America vol. 106,38 (2009): 16233-8. doi:10.1073/pnas.0908301106 [3] Koruza, Katarina et al. “Biophysical Characterization of Cancer-Related Carbonic Anhydrase IX.” International journal of molecular sciences vol. 21,15 5277. 25 Jul. 2020, doi:10.3390/ijms21155277 [4] Alterio, Vincenzo et al. “Multiple binding modes of inhibitors to carbonic anhydrases: how to design specific drugs targeting 15 different isoforms?.” Chemical reviews vol. 112,8 (2012): 4421-68. doi:10.1021/cr200176r ER -
RAVASZOVÁ, Karin, Alena HOFROVÁ a Jozef HRITZ. Preparation of catalytic domain of carbonic anhydrase IX with different protein tags. In \textit{XXII. Setkání biochemiků a molekulárních biologů}. 2022.
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