title: Real-time monitoring of disintegration activity of catalytic core domain of HIV-1 integrase using molecular beacon.
abstract: HIV-1 integrase, an essential enzyme for retroviral replication, is a validated target for anti-HIV therapy development. The catalytic core domain of integrase (IN-CCD) is capable of catalyzing disintegration reaction. In this work, a hairpin-shaped disintegration substrate was designed and validated by enzyme-linked immunosorbent assay; a molecular beacon-based assay was developed for disintegration reaction of IN-CCD. Results showed that the disintegration substrate could be recognized and catalyzed by IN-CCD, and the disintegration reaction can be monitored according to the increase of fluorescent signal. The assay can be applied to real-time detection of disintegration with advantages of simplicity, high sensitivity, and excellent specificity.
authors.affiliation: Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, People's Republic of China.
authors.forename: Da-wei
authors.initials: DW
authors.lastname: Zhang
authors.forename: Ming-ming
authors.initials: MM
authors.lastname: Zhao
authors.forename: Hong-qiu
authors.initials: HQ
authors.lastname: He
authors.forename: Shun-xing
authors.initials: SX
authors.lastname: Guo
mesh_terms: D001483:Base Sequence; D020134:Catalytic Domain; D057075:Enzyme Assays / Q000379:methods*; D019427:HIV Integrase / Q000737:chemistry* / Q000378:metabolism*; D015497:HIV-1 / Q000201:enzymology*; D015345:Oligonucleotide Probes / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors
chemical_list: D015345:Oligonucleotide Probes; D019427:HIV Integrase
keywords: Disintegration; HIV-1; IN-CCD; Integrase; Molecular beacon