Purification of the Xenopus laevis double-stranded RNA adenosine deaminase
- PMID: 8144588
Purification of the Xenopus laevis double-stranded RNA adenosine deaminase
Abstract
A double-stranded RNA adenosine deaminase that catalyzes the conversion of adenosines to inosines in duplex RNA substrates was purified to near homogeneity from Xenopus laevis eggs. The final specific activity was approximately 2.0 nmol of inosine min-1 mg-1 at 25 degrees C and pH 7.9 with a 794-base pair RNA substrate. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a single major approximately 120-kDa protein band by silver staining. The purified enzyme migrated with an apparent molecular mass of 90 +/- 10 kDa during high performance liquid chromatography. Gel filtration of the partially purified enzyme gave an apparent molecular mass of 210 +/- 20 kDa, suggesting that the enzyme may dimerize or associate with other cellular components. Substrate modification was inhibited by excess substrate, thiol reagents, heparin, and moderate concentrations of monovalent cations.
Similar articles
-
Purification and properties of double-stranded RNA-specific adenosine deaminase from calf thymus.Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10596-600. doi: 10.1073/pnas.91.22.10596. Proc Natl Acad Sci U S A. 1994. PMID: 7937998 Free PMC article.
-
Purification and characterization of double-stranded RNA adenosine deaminase from bovine nuclear extracts.J Biol Chem. 1994 May 6;269(18):13480-9. J Biol Chem. 1994. PMID: 8175781
-
Purification of native and recombinant double-stranded RNA-specific adenosine deaminases.Methods. 1998 May;15(1):51-62. doi: 10.1006/meth.1998.0605. Methods. 1998. PMID: 9614652
-
Double-stranded RNA adenosine deaminase as a potential mammalian RNA editing factor.Semin Cell Biol. 1993 Aug;4(4):285-93. doi: 10.1006/scel.1993.1034. Semin Cell Biol. 1993. PMID: 8241470 Review.
-
Double-stranded RNA binding proteins and their substrates.Nucleic Acids Symp Ser. 1995;(33):13-5. Nucleic Acids Symp Ser. 1995. PMID: 8643348 Review.
Cited by
-
Hepatitis D virus RNA editing: specific modification of adenosine in the antigenomic RNA.J Virol. 1995 Dec;69(12):7593-600. doi: 10.1128/JVI.69.12.7593-7600.1995. J Virol. 1995. PMID: 7494266 Free PMC article.
-
Purification and properties of double-stranded RNA-specific adenosine deaminase from calf thymus.Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10596-600. doi: 10.1073/pnas.91.22.10596. Proc Natl Acad Sci U S A. 1994. PMID: 7937998 Free PMC article.
-
ADAR1 and ADAR2 expression and editing activity during forebrain development.Dev Neurosci. 2009;31(3):223-37. doi: 10.1159/000210185. Epub 2009 Mar 27. Dev Neurosci. 2009. PMID: 19325227 Free PMC article.
-
Abnormalities in A-to-I RNA editing patterns in CNS injuries correlate with dynamic changes in cell type composition.Sci Rep. 2017 Mar 7;7:43421. doi: 10.1038/srep43421. Sci Rep. 2017. PMID: 28266523 Free PMC article.
-
An ADAR that edits transcripts encoding ion channel subunits functions as a dimer.EMBO J. 2003 Jul 1;22(13):3421-30. doi: 10.1093/emboj/cdg327. EMBO J. 2003. PMID: 12840004 Free PMC article.
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Other Literature Sources
Research Materials
Miscellaneous
