3DLK

Crystal Structure of an engineered form of the HIV-1 Reverse Transcriptase, RT69A

Summary for 3DLK

• Entry DOI: 10.2210/pdb3dlk/pdb
• Descriptor: Reverse transcriptase/ribonuclease H, p51 RT, SULFATE ION, ... (4 entities in total)
• Functional Keywords: aids, hiv, reverse transcriptase, rt, crystal engineering, rna-binding, rna-directed dna polymerase, transferase, viral nucleoprotein
• Biological source: Human immunodeficiency virus type 1 BH10 (HIV-1); More
• Cellular location: Gag-Pol polyprotein: Host cell membrane; Lipid-anchor. Matrix protein p17: Virion membrane; Lipid- anchor . Capsid protein p24: Virion . Nucleocapsid protein p7: Virion . Reverse transcriptase/ribonuclease H: Virion . Integrase: Virion : P03366 P03366
• Total number of polymer chains: 2
• Total formula weight: 113574.14

Authors

Ho, W.C.,Bauman, J.D.,Himmel, D.M.,Das, K.,Arnold, E. (deposition date: 2008-06-27, release date: 2008-10-07, Last modification date: 2023-08-30)

Primary citation

Bauman, J.D.,Das, K.,Ho, W.C.,Baweja, M.,Himmel, D.M.,Clark, A.D.,Oren, D.A.,Boyer, P.L.,Hughes, S.H.,Shatkin, A.J.,Arnold, E.
Crystal engineering of HIV-1 reverse transcriptase for structure-based drug design.
Nucleic Acids Res., 36:5083-5092, 2008

PubMed Abstract: HIV-1 reverse transcriptase (RT) is a primary target for anti-AIDS drugs. Structures of HIV-1 RT, usually determined at approximately 2.5-3.0 A resolution, are important for understanding enzyme function and mechanisms of drug resistance in addition to being helpful in the design of RT inhibitors. Despite hundreds of attempts, it was not possible to obtain the structure of a complex of HIV-1 RT with TMC278, a nonnucleoside RT inhibitor (NNRTI) in advanced clinical trials. A systematic and iterative protein crystal engineering approach was developed to optimize RT for obtaining crystals in complexes with TMC278 and other NNRTIs that diffract X-rays to 1.8 A resolution. Another form of engineered RT was optimized to produce a high-resolution apo-RT crystal form, reported here at 1.85 A resolution, with a distinct RT conformation. Engineered RTs were mutagenized using a new, flexible and cost effective method called methylated overlap-extension ligation independent cloning. Our analysis suggests that reducing the solvent content, increasing lattice contacts, and stabilizing the internal low-energy conformations of RT are critical for the growth of crystals that diffract to high resolution. The new RTs enable rapid crystallization and yield high-resolution structures that are useful in designing/developing new anti-AIDS drugs.

PubMed: 18676450
DOI: 10.1093/nar/gkn464

Experimental method

X-RAY DIFFRACTION (1.85 Å)