MAGE (melanoma-associated antigen) Domain structural studies using NMR

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Authors

BATHULA Sreenivas Reddy PALEČEK Jan ŽÍDEK Lukáš SKLENÁŘ Vladimír

Year of publication 2009
Type Appeared in Conference without Proceedings
MU Faculty or unit

Faculty of Science

Citation
Description MAGE (melanoma-associated antigen) gene family is huge and widely spread over different types of species. These proteins importance rose because of their high expression in tumor cells together with their antigen presenting nature. In human, more than 30 proteins have this domain. In some MAGE family members MAGE domain repeats. However, exact structure or function of these proteins is not clearly known. Proteins containing MHD show interactions with various important cellular proteins. Expected MAGE domain function is closely related to cell development, DNA Repair and apoptosis. Using 2 liters of 15N isotopically labeled media, we produced 10 mg/ml MAGE A4 protein (MAGE Homology Domain, 25 kDa ). It corresponds to 700ul, 0.56 mM protein sample in 25 mM phosphate buffer in normal saline (136-150 mM NaCl), pH 7.32. When we run HSQC measurements on 600 MHz NMR system (equipped with TCI cryoprobehead, 16 scans at 30degC), we obtained the spectra where peaks were well resolved and number of peaks was close to the expected number of peaks based on the amino acid sequence.
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