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

BATHULA, Sreenivas Reddy, Jan PALEČEK, Lukáš ŽÍDEK and Vladimír SKLENÁŘ. MAGE (melanoma-associated antigen) Domain structural studies using NMR. In Central European NMR Meeting, Valtice, CR. 2009.

Basic information

• Original name: MAGE (melanoma-associated antigen) Domain structural studies using NMR
• Name in Czech: Studium MAGE domény pomocí NMR spektroskopie
• Authors: BATHULA, Sreenivas Reddy, Jan PALEČEK, Lukáš ŽÍDEK and Vladimír SKLENÁŘ.
• Edition: Central European NMR Meeting, Valtice, CR, 2009.

Other information

• Original language: English
• Type of outcome: Presentations at conferences
• Field of Study: 10600 1.6 Biological sciences
• Country of publisher: Czech Republic
• Confidentiality degree: is not subject to a state or trade secret
• Organization unit: Faculty of Science
• Keywords in English: melanoma antigen MAGE protein NMR

Abstract

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.

Abstract (in Czech)

Rodina genů zvaných MAGE (melanoma-associated antigen)je široce rozšířená v mnoha organismech. Hladina exprese těchto genů je zvýšená u nádorových buněk společně s výskytem jejich fragmentů jako antigenních agens. V lidských buňkách je více jak 30 proteinů obsahujících MAGE doménu (MHD). Přesná struktura této domény není známá. Naprodukovali jsme (ze 2 litrů 15N isotop media) 10 mg/ml MAGE A4 proteinu (25 kDa), což odpovídá v 700ul koncentraci 0.56 mM proteinu. Měřili jsme HSQC na 600 MHz NMR systemu (TCI cryoprobehead, 16 scans při 30degC), a získali jsme spektra s dobře rozlišenými píky.

Links

• IAA501630902, research and development project: Name: Charakterizace MAGE proteinů: kofaktory E3 ubiquitin ligáz?

Investor: Academy of Sciences of the Czech Republic

• MSM0021622415, plan (intention): Name: Molekulární podstata buněčných a tkáňových regulací

Investor: Ministry of Education, Youth and Sports of the CR, Molecular basis of cell and tissue regulations