Premium
A directed approach to improving the solubility of Moloney murine leukemia virus reverse transcriptase
Author(s) -
Das Debanu,
Georgiadis Millie M.
Publication year - 2001
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.16301
Subject(s) - reverse transcriptase , murine leukemia virus , retrovirus , solubility , enzyme , dna , chemistry , rna , polymerase , microbiology and biotechnology , virus , biology , virology , biochemistry , gene , organic chemistry
One of the difficulties that can impede structural work on a molecule of interest is limited solubility. Although functionally similar to the human immunodeficiency virus type‐1 reverse transcriptase (HIV‐1 RT), the Moloney murine leukemia virus reverse transcriptase (MMLV RT) differs both in architecture and solubility properties. Reverse transcriptase is an essential retroviral enzyme that replicates the single‐stranded RNA genome of the retrovirus producing a double‐stranded DNA copy, which is subsequently integrated into the host's genome. We have introduced a single amino acid substitution in the connection domain of an N‐terminally truncated MMLV RT (L435K) that significantly improves the solubility of the enzyme eliminating the need for nonionic detergents in buffering storage solutions. The substituted enzyme retains near wild‐type polymerase activity. An important consequence of the improved solubility of the L435K MMLV RT has been the ability to obtain diffraction quality crystals.