In vitro translation to study HIV protease activity

Zene Matsuda, Mutsunori Murahashi, Kosuke Miyauchi, Jun Komano, Kazuhiro Morishita, Akihiko Okayama, Hirohito Tsubouchi

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Citations (Scopus)

Abstract

HIV-1 is an etiological agent of AIDS. One of the targets of the current anti-HIV-1 combination chemotherapy, called highly active antiretroviral therapy (HAART), is HIV-1 protease (PR), which is responsible for the processing of viral structural proteins and, therefore, essential for virus replication. Here, we describe an in vitro transcription/translation-based method of phenotyping HIV-1 PR. In this system, both substrate and PR for the assay can be prepared by in vitro transcription/translation. Protease activity is estimated by the cleavage of a substrate, as measured by enzyme-linked immunosorbent assay (ELISA). This assay is safe, rapid, and requires no special facility to be carried out. Our rapid phenotyping method of HIV-1 PR may help evaluate drug resistance, useful when choosing an appropriate therapeutic regiment, and could potentially facilitate the discovery of new drugs effective against HIV-1 PR.

Original languageEnglish
Title of host publicationIn Vitro Transcription and Translation Protocols
Subtitle of host publicationSecond Edition
Pages135-149
Number of pages15
Volume375
DOIs
Publication statusPublished - Apr 4 2007
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume375
ISSN (Print)1064-3745

Fingerprint

HIV Protease
HIV-1
Peptide Hydrolases
Viral Structural Proteins
Highly Active Antiretroviral Therapy
Drug Discovery
Virus Replication
Combination Drug Therapy
Drug Resistance
Acquired Immunodeficiency Syndrome
Enzyme-Linked Immunosorbent Assay
Human immunodeficiency virus 1 p16 protease
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

Matsuda, Z., Murahashi, M., Miyauchi, K., Komano, J., Morishita, K., Okayama, A., & Tsubouchi, H. (2007). In vitro translation to study HIV protease activity. In In Vitro Transcription and Translation Protocols: Second Edition (Vol. 375, pp. 135-149). (Methods in Molecular Biology; Vol. 375). https://doi.org/10.1385/1-59745-388-9:135

In vitro translation to study HIV protease activity. / Matsuda, Zene; Murahashi, Mutsunori; Miyauchi, Kosuke; Komano, Jun; Morishita, Kazuhiro; Okayama, Akihiko; Tsubouchi, Hirohito.

In Vitro Transcription and Translation Protocols: Second Edition. Vol. 375 2007. p. 135-149 (Methods in Molecular Biology; Vol. 375).

Research output: Chapter in Book/Report/Conference proceedingChapter

Matsuda, Z, Murahashi, M, Miyauchi, K, Komano, J, Morishita, K, Okayama, A & Tsubouchi, H 2007, In vitro translation to study HIV protease activity. in In Vitro Transcription and Translation Protocols: Second Edition. vol. 375, Methods in Molecular Biology, vol. 375, pp. 135-149. https://doi.org/10.1385/1-59745-388-9:135
Matsuda Z, Murahashi M, Miyauchi K, Komano J, Morishita K, Okayama A et al. In vitro translation to study HIV protease activity. In In Vitro Transcription and Translation Protocols: Second Edition. Vol. 375. 2007. p. 135-149. (Methods in Molecular Biology). https://doi.org/10.1385/1-59745-388-9:135
Matsuda, Zene ; Murahashi, Mutsunori ; Miyauchi, Kosuke ; Komano, Jun ; Morishita, Kazuhiro ; Okayama, Akihiko ; Tsubouchi, Hirohito. / In vitro translation to study HIV protease activity. In Vitro Transcription and Translation Protocols: Second Edition. Vol. 375 2007. pp. 135-149 (Methods in Molecular Biology).
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