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Journal : Journal of Microbial Systematics and Biotechnology

Development of a dimer-based screening system for dimerization inhibitor of HIV-1 protease I Dewa Agung Panji Dwipayana; Yana Maolana Syah; Reza Aditama; Feraliana Feraliana; Azzania Fibriani
Journal of Microbial Systematics and Biotechnology Vol 2, No 2 (2020): December 2020
Publisher : Microbiology Division, Research Center for Biology, Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.37604/jmsb.v2i2.42

Abstract

An in vitro dimer-based screening system (DBSS) for selecting new HIV-1 protease dimerization inhibitor candidates from natural compounds had been established. This system utilizes a fusion between HIV-1 protease and dimer binding domain of AraC protein (proteaseHIV1-AraCDBD) where fluorescence signal will be emitted in the presence of HIV-1 protease inhibitor. However, this screening system had not been evaluated. Therefore, this study was aimed to evaluate it in recombinant Escherichia coli culture. The expression of proteaseHIV1-AraCDBD fusion gene was observed for 18 hours. Its crude lysate isolation was done once every 3 hours and analyzed using SDS PAGE. To test the DBSS, darunavir was used as positive control, and Nigella sativa extract (JH3) was used as the test compound. The results of SDS PAGE analysis on crude lysates presented a ~24.2 kDa band, which was the predicted size of the proteaseHIV1-AraCDBD fusion protein based on its amino acid sequence. The growth curve and protein expression profiles revealed that the 15 hours was the optimum culture age to be used in the screening system. Darunavir testing in DBSS showed an increase in fluorescence signal compared to the negative control. The same increase in fluorescence signal was also obtained from the JH3 compound test. In conclusion, DBSS could be used as an assay to screen for new HIV-1 protease inhibitors, and the JH3 compound demonstrated the ability to inhibit HIV-1 protease dimerization.
Co-Authors A.A. Ketut Agung Cahyawan W Achmad Saifuddin Noer Affrida Affrida Aisyah Aisyah Aisyah Aisyah Alan Jefferson Aliefman Hakim Asep Aripin Asep Kadarohman Azzania Fibriani Bambang Ariwahjoedi Buchari Buchari Buchari Buchari Bunbun Bundjali Deana Wahyuningrum Debbie Soefie Soefie Dessy Natalia Devitri Amisa Dian Nugraheni Didin - Mujahidin Didin Mujahidin Didin Mujahidin Didin Mujahidin Didin Mujahidin Didin Mujahidin Didin Mujahidin Didin Mujahidin Dikhi Firmansyah Eliza Eliza Elvira Hermawati Elvira Hermawati Emilia L Ghisalberti Emilia L. Ghisalberti Emilio L Ghisalberti Endang Rosdiana Eri Bachtiar Eri Bakhtiar Euis Holishotan Hakim Euis Holisotan Hakim Euis Holisotan Hakim Euis Holisotan Hakim Euis Holisotan Hakim Euis Holisotan Hakim Euis Holisotan Hakim Euis Holisotan Hakim Euis Holisotan Hakim Euis Holisotasn Hakim Feraliana Feraliana Ferlinahayati Ferlinahayati Firdaus, Firdaus Hanapi Usman Hartiwi Diastuti Hartiwi Diastuti Hartiwi Diastuti Haryoto Haryoto Hendra Helmanto Hernandi Sujono Hiromitsu Takayama Hiromitsu Takayama I Dewa Agung Panji Dwipayana I Nyoman Adi Winata Ihsanawati Ihsanawati Ikram M. Said Ilim Ilim Ilimu, Edi Iqbal M. Choudhary Jalifah Latip Jalifah Latip Joko Santoso Kholifatu Rosyidah Laily Bin Din Lia Dewi Juliawati Lia Dewi Juliawati Lia Dewi Juliawaty Lia Dewi Juliawaty Lia Dewi Juliawaty Liliasari Liliasari Lukman Makmur Lukman Makmur Lukman Makmur Lukman Makmur Lukman Makmur Lukman Makmur M I Choudhary Marc Jeannin Mariko Katajima Mariko Kitajima Mariko Kitajima Marlia Singgih Marlia Singgih Marlia Singgih Masatake Niwa Melina Melina Muhamad Abdulkadir Martoprawiro Muhammad N. Jalaluddin Muhtadi Muhtadi Nanik Siti Aminah Nia Sri Hardianti Nila Berghuis Tamaela Nila Berghuis Tamaela, Nila Berghuis Nizar Happyana Nizar Happyana Nordin Hj Lajis Norio Aimi Norio Aimi Norio Aimi Noviany Noviany Nunuk H Soekamto Nunuk Hariani Soekamto Purkan Purkan Reza Aditama Rusjdi Tamin Sadijah Achmad Sahidin . Sahidin Sahidin Senadi Budiman Siti Nurbayti Siti Zubaidah Sjamsul Arifin Achmad Sjamsul Arifin Achmad Sjamsul Arifin Achmad Sjamsul Arifin Achmad Sjamsul Arifin Achmad Sjamsul Arifin Achmad Sjamsul Arifin Achmad Sjamsul Arifin Achmad Soerya Dewi Marliyana Sri Atun Suci Zulaikha Hildayani Sukrido Sukrido Suwandri Suwandri Suwandri Suwandri Tia Setiawati Tjodi Harlim Tri Cahyanto Valentina Adimurti Valentina Adimurti Kusumaningtyas Wasinton Simanjuntak Windayani, Neneng Yaya Rukayadi Yuliana, Trisna Zakaria Zakaria