Purification, Characterization and Crystallization of Acanthamoeba Profilin Expressed in Escherichia coli

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 profilin from Acanthamoeba -based on fluorescence monitored urea denaturation, circular dichroism, actin-nucleotide e...

Ausführliche Beschreibung

Gespeichert in:

Bibliographische Detailangaben
Veröffentlicht in:	Journal of molecular biology 1994-02, Vol.236 (3), p.950-952
Hauptverfasser:	Almo, S.C., Pollard, T.D., Way, M., Lattman, E.E.
Format:	Artikel
Sprache:	eng
Schlagworte:	Acanthamoeba - metabolism Acanthamoeba castellanii Actins - chemistry Actins - metabolism Analytical, structural and metabolic biochemistry Animals Binding and carrier proteins Biological and medical sciences Circular Dichroism Contractile Proteins - chemistry Contractile Proteins - isolation & purification Crystalline structure Crystallization Crystallography, X-Ray cytoskeleton Escherichia coli expression Fundamental and applied biological sciences. Psychology Kinetics Microfilament Proteins - chemistry Microfilament Proteins - isolation & purification Microfilament Proteins - metabolism Molecular biophysics Muscles - metabolism profilin Profilins Protein Conformation Protein Denaturation protein structure Proteins Rabbits Recombinant Proteins - chemistry Recombinant Proteins - isolation & purification Recombinant Proteins - metabolism Structure in molecular biology
Online-Zugang:	Volltext
Tags:	Tag hinzufügen Keine Tags, Fügen Sie den ersten Tag hinzu!

Beschreibung
Zusammenfassung:	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 profilin from Acanthamoeba -based on fluorescence monitored urea denaturation, circular dichroism, actin-nucleotide exchange rate and the K d 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, β=109·6°. These crystals contain one molecule in the asymmetric unit and diffract to 2·0 Å.
ISSN:	0022-2836 1089-8638
DOI:	10.1006/jmbi.1994.1200