2je4
From Proteopedia
|
ATOMIC-RESOLUTION CRYSTAL STRUCTURE OF CHEMICALLY-SYNTHESIZED HIV-1 PROTEASE IN COMPLEX WITH JG-365
Overview
As part of our ongoing studies of the human immunodeficiency virus type 1, (HIV-1) protease enzyme, we set out to develop a modular chemical, synthesis of the protein from multiple peptide segments. Our initial, attempts were frustrated by the insolubility of intermediate peptide, products. To overcome this problem, we designed a synthetic strategy, combining the solubility-enhancing properties of C-terminal (Arg)n tags, and the biological phenomenon of autoprocessing of the Gag-Pol polyprotein, that occurs during maturation of the HIV-1 virus in vivo. Synthesis of a, 119-residue peptide chain containing 10 residues of the reverse, transcriptase (RT) open reading frame plus an (Arg)10 tag at the, C-terminus was straightforward by native chemical ligation followed by, conversion of the Cys residues to Ala by Raney nickel desulfurization. The, product polypeptide itself completed the final synthetic step by removing, the C-terminal modification under folding conditions, to give the mature, 99-residue polypeptide. High-purity homodimeric HIV-1 protease protein was, obtained in excellent yield and had full enzymatic activity; the structure, of the synthetic enzyme was confirmed by X-ray crystallography to a, resolution of 1.07 A. This efficient modular synthesis by a biomimetic, autoprocessing strategy will enable the facile synthesis of unique, chemical analogues of the HIV-1 protease to further elucidate the, molecular basis of enzyme catalysis.
About this Structure
2JE4 is a Protein complex structure of sequences from Human immunodeficiency virus 1 with ACT, SO4 and GOL as ligands. Structure known Active Site: AC1. Full crystallographic information is available from OCA.
Reference
Modular Total Chemical Synthesis of a Human Immunodeficiency Virus Type 1 Protease., Johnson EC, Malito E, Shen Y, Rich D, Tang WJ, Kent SB, J Am Chem Soc. 2007 Sep 19;129(37):11480-11490. Epub 2007 Aug 18. PMID:17705484
Page seeded by OCA on Mon Nov 5 13:02:29 2007
