A directed approach to improving the solubility of Moloney murine leukemia virus reverse transcriptase

Debanu Das, Millie Georgiadis

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1936-1941
Number of pages6
JournalProtein Science
Volume10
Issue number10
DOIs
StatePublished - 2001
Externally publishedYes

Fingerprint

Moloney murine leukemia virus
RNA-Directed DNA Polymerase
Viruses
Solubility
Enzymes
Genes
Genome
Retroviridae
Amino Acid Substitution
Detergents
HIV-1
Substitution reactions
Diffraction
RNA
Amino Acids
Crystals
Molecules
DNA

Keywords

  • Crystallization
  • Moloney murine leukemia virus
  • Mutant
  • Reverse transcriptase
  • Solubility

ASJC Scopus subject areas

  • Biochemistry

Cite this

A directed approach to improving the solubility of Moloney murine leukemia virus reverse transcriptase. / Das, Debanu; Georgiadis, Millie.

In: Protein Science, Vol. 10, No. 10, 2001, p. 1936-1941.

Research output: Contribution to journalArticle

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