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PMID: 2217189 Published · ppublish English Journal Article

Purification and characterization of recombinant Rev protein of human immunodeficiency virus type 1.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 87 ·No. 19 ·1990-10-00 ·Pages 7593-7

Nalin CM, Purcell RD, Antelman D, Mueller D, Tomchak L, Wegrzynski B, McCarney E, Toome V, Kramer R, Hsu MC

Abstract

Recombinant Rev protein of human immunodeficiency virus type 1 has been expressed in Escherichia coli and purified by ion-exchange and gel-filtration chromatography. Specific binding of the purified protein to the Rev-responsive element of the viral RNA is demonstrated. Physical characterization of the purified protein by circular dichroism and intrinsic fluorescence spectroscopy indicate that the protein preparation is suitable for structural analysis. Circular dichroism measurements show that the protein is approximately 40-45% alpha-helix. Tryptophan fluorescence measurements suggest that the single tryptophan residue is located near the surface of the protein. Gel-filtration chromatography of the protein indicates that it has an apparent molecular mass of 33,000 daltons. This suggests that the protein in solution forms a stable tetramer consisting of monomers having molecular mass of 13,000 daltons.

Related Genes
rev
MeSH Terms
Chromatography, Gel Circular Dichroism Cloning, Molecular Escherichia coli/genetics Gene Products, rev/genetics,isolation & purification,metabolism Genes, Synthetic HIV-1/genetics,metabolism Kinetics Plasmids Protein Binding Protein Conformation Recombinant Proteins/isolation & purification Spectrometry, Fluorescence Spectrophotometry, Ultraviolet rev Gene Products, Human Immunodeficiency Virus
Chemicals
Gene Products, rev Recombinant Proteins rev Gene Products, Human Immunodeficiency Virus
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Nalin C M
Department of Protein Biochemistry, Roche Research Center, Hoffmann-La Roche Inc., Nutley, NJ 07110.
Purcell R D
Antelman D
Mueller D
Tomchak L
Wegrzynski B
McCarney E
Toome V
Kramer R
Hsu M C
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.87.19.7593
Published
1990-10-00
Pages
7593-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC54794
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.87.19.7593 PMC Full Text View journal details
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