Purification characterization and crystallization of Acanthamoeba profilin expressed in Escherichia coli

S. C. Almo; T. D. Pollard; M. Way; E. E. Lattman

doi:10.1006/jmbi.1994.1200

Purification characterization and crystallization of Acanthamoeba profilin expressed in Escherichia coli

S. C. Almo, T. D. Pollard, M. Way, E. E. Lattman

Research output: Contribution to journal › Article › peer-review

31 Scopus citations

Abstract

Profilin (isoform I) from Acanthamoeba castellani was expressed in Escherichia coli using a bacteriophage T7-based expression vector. The recombinant material is similar to authentic profilm from Acanthamoeba-based on fluorescence monitored urea denaturation, circular dichroisrn, actin-nuoleotide exchange rate and the K_a for rabbit skeletal actin. This recombinant material crystallized from 80% saturated sodium potassium tartrate, yielding monoclinic crystals, space group C2, a = 91·4 Å, b =37·4 Å, c = 34·7 Å, α =109·6°. These crystals contain one molecule in the asymmetric unit and diffract to 2·0 Å.

Original language	English (US)
Pages (from-to)	950-952
Number of pages	3
Journal	Journal of Molecular Biology
Volume	236
Issue number	3
DOIs	https://doi.org/10.1006/jmbi.1994.1200
State	Published - Feb 24 1994
Externally published	Yes

Keywords

Crystallization
Cytoskeleton
Expression
Profilin
Protein structure

ASJC Scopus subject areas

Biophysics
Structural Biology
Molecular Biology

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10.1006/jmbi.1994.1200

Cite this

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title = "Purification characterization and crystallization of Acanthamoeba profilin expressed in Escherichia coli",

abstract = "Profilin (isoform I) from Acanthamoeba castellani was expressed in Escherichia coli using a bacteriophage T7-based expression vector. The recombinant material is similar to authentic profilm from Acanthamoeba-based on fluorescence monitored urea denaturation, circular dichroisrn, actin-nuoleotide exchange rate and the Ka for rabbit skeletal actin. This recombinant material crystallized from 80% saturated sodium potassium tartrate, yielding monoclinic crystals, space group C2, a = 91·4 {\AA}, b =37·4 {\AA}, c = 34·7 {\AA}, α =109·6°. These crystals contain one molecule in the asymmetric unit and diffract to 2·0 {\AA}.",

keywords = "Crystallization, Cytoskeleton, Expression, Profilin, Protein structure",

author = "Almo, {S. C.} and Pollard, {T. D.} and M. Way and Lattman, {E. E.}",

year = "1994",

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doi = "10.1006/jmbi.1994.1200",

language = "English (US)",

volume = "236",

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journal = "Journal of Molecular Biology",

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TY - JOUR

T1 - Purification characterization and crystallization of Acanthamoeba profilin expressed in Escherichia coli

AU - Almo, S. C.

AU - Pollard, T. D.

AU - Way, M.

AU - Lattman, E. E.

PY - 1994/2/24

Y1 - 1994/2/24

N2 - Profilin (isoform I) from Acanthamoeba castellani was expressed in Escherichia coli using a bacteriophage T7-based expression vector. The recombinant material is similar to authentic profilm from Acanthamoeba-based on fluorescence monitored urea denaturation, circular dichroisrn, actin-nuoleotide exchange rate and the Ka for rabbit skeletal actin. This recombinant material crystallized from 80% saturated sodium potassium tartrate, yielding monoclinic crystals, space group C2, a = 91·4 Å, b =37·4 Å, c = 34·7 Å, α =109·6°. These crystals contain one molecule in the asymmetric unit and diffract to 2·0 Å.

AB - Profilin (isoform I) from Acanthamoeba castellani was expressed in Escherichia coli using a bacteriophage T7-based expression vector. The recombinant material is similar to authentic profilm from Acanthamoeba-based on fluorescence monitored urea denaturation, circular dichroisrn, actin-nuoleotide exchange rate and the Ka for rabbit skeletal actin. This recombinant material crystallized from 80% saturated sodium potassium tartrate, yielding monoclinic crystals, space group C2, a = 91·4 Å, b =37·4 Å, c = 34·7 Å, α =109·6°. These crystals contain one molecule in the asymmetric unit and diffract to 2·0 Å.

KW - Crystallization

KW - Cytoskeleton

KW - Expression

KW - Profilin

KW - Protein structure

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DO - 10.1006/jmbi.1994.1200

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JF - Journal of Molecular Biology

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Purification characterization and crystallization of Acanthamoeba profilin expressed in Escherichia coli

Abstract

Keywords

ASJC Scopus subject areas

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