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| Type |
Oral Presentation |
| Area |
Oral Presentation of Young Analytical Chemists Ⅰ |
| Room No. |
303호 |
| Time |
THU 09:34-: |
| Code |
ANAL1.O-10 |
| Subject |
NMR study of Arginine ε-NH NMR Signal Assignment in Human Cytosolic Adenylate Kinase 1
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| Authors |
김길훈, 원호식*
한양대학교 응용화학과, Korea
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| Abstract |
In many of nucleotide binding proteins, Arg residues are known to play important roles in substrate binding through the electrostatic interaction of positive charge on the Arg side chain with the negative charge of phosphoryl group. Adenylate kinase1(AK1) has some Arg residues necessary for binding its substrates, MgATP, AMP. To investigate the interactions between Arg side chains and the substrate in aqueous solution, we observed the signals of Arg ε-NHs of 15N-labeled AK using 1H, 15N two dimensional NMR. The experiments with HSQC pulse sequence detected 13 signals corresponding to all Arg residues on human cytosolic AK1(hAK1). In NMR spectra, we used single quantum coherence SQC as pulse sequence, only the ε-NH signal on Arg side chain can be detected in the specific resonance region. These signals are good markers to study the inter actions between the substrate and proteins with high molecular weight. However, this process is often hard to assign near the terminus of the long side chain such as Arg residues. Therefore, we focused of on six conserved Arg and assigned only these signals by comparison with the spectra of each Arg mutant. The 6 signals derived from 6 Arg residues conserve beyond species (Arg[44], [97], [128], [132], [138], and [149]) were assigned using each mutants which substituted by Ala residue (R44A, R97A, R128A, R132A, R138A and R149A). This method had the problem that the structural perturbation induced by the substitution of amino acid would make the assignment impossible. In the spectra of R44A, R97A, and R128A mutants, there were few problems. Although some perturbations appeared by occurred in R132A, R138A, and R149A of AK1 mutants, the consideration of the assignment for the other signals permitted these signal to be assigned. |
| E-mail |
kghpotter@naver.com |
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119th General Meeting of the KCS