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 exchange rate and the Kd for rabbit skeletal actin. This recombinant material crystallized from 80% saturated sodium potassium tartrate, yielding monoclinic crystals, space group C2, a = 91.4 A, b = 37.4 A, c = 34.7 A, beta = 109.6 degrees. These crystals contain one molecule in the asymmetric unit and diffract to 2.0 A.
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Department of Cell Biology and Anatomy, Johns Hopkins School of Medicine, Baltimore, MD 21205.
Pollard
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Way
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Lattman
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D000048:Acanthamoeba / Q000378:metabolism*; D000199:Actins / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D002942:Circular Dichroism; D003285:Contractile Proteins / Q000737:chemistry / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli; D007700:Kinetics; D008840:Microfilament Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D009132:Muscles / Q000378:metabolism; D051304:Profilins; D011487:Protein Conformation; D011489:Protein Denaturation; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
D000199:Actins; D003285:Contractile Proteins; D008840:Microfilament Proteins; D051304:Profilins; D011994:Recombinant Proteins