期刊
JOURNAL OF CHEMICAL EDUCATION
卷 94, 期 10, 页码 1547-1551出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jchemed.7b00160
关键词
Heterocycles; Nucleic Acids/DNA/RNA; Spectroscopy; Graduate Education/Research; Biochemistry; Interdisciplinary/Multidisciplinary; Hands-On Learning/Manipulatives; Laboratory Instruction; Bioanalytical Chemistry; Upper-Division Undergraduate
资金
- FCT-Foundation for Science and Technology [SFRH/BD/122953/2016, UID/Multi/00709/2013]
- Fundo Social Europeu [IF/00959/2015]
- FEDER funds through the POCI-COMPETE-Operational Programme Competitiveness and Internationalisation in Axis I-Strengthening research, technological development and innovation [POCI-01-0145-FEDER-007491]
- Programa Operacional Potencial Humano
- Fundação para a Ciência e a Tecnologia [SFRH/BD/122953/2016] Funding Source: FCT
Circular dichroism (CD) has emerged as one of the standard biophysical techniques for the study of guanine-quadruplex (G4) folding, cation effect, and ligand binding. The utility of this technique is based on its robustness, ease of use, and requirement of only small quantities of nucleic acid. This experiment is also extendable to the classroom and adaptable to students from graduate school in a multidisciplinary area, bridging chemistry, biochemistry, and medicinal chemistry. Specifically, students employ a solution of a nucleic acid that can adopt a G4 conformation, anneal it in the presence of a metal cation, and assess the folding and topology by the characteristic CD signatures. During the class experiment, it was also possible to interconvert the G4 topology by changing the cation type, and to determine the melting temperature of the G4 structure in the absence and presence of molecules called G4 ligands. The current experiment used the commercially available porphyrin TMPyP4 as a G4 ligand and human telomeric 22AG and mutated oncogene promoter c-MYC G4 structures to illustrate the CD applicability in a context that can be explored or adapted to a student's course level and topic.
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