Overproduction in Escherichia coli and purification of the hemolytic protein sticholysin II from the sea anemone Stichodactyla helianthus.
The cDNA coding for the cytolytic toxins sticholysin I and sticholysin II from the sea anemone Stichodactyla helianthus has been isolated, cloned in pUC18, and sequenced. A 6His-tagged version of sticholysin II has been overproduced in Escherichia coli and purified to homogeneity in milligram amounts. Conformational and functional analyses of recombinant sticholysin II do not reveal any significant difference when compared to the natural cytolysin.
de los Ríos
V
V
Departamento de Bioquímica y Biología Molecular, Facultad de Química, Universidad Complutense, Madrid, 28040, Spain.
Oñaderra
M
M
Martínez-Ruiz
A
A
Lacadena
J
J
Mancheño
J M
JM
Martínez del Pozo
A
A
Gavilanes
J G
JG
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D003064:Cnidarian Venoms / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D006460:Hemolysin Proteins / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009930:Organic Chemicals; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D012615:Sea Anemones / Q000235:genetics* / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
D003064:Cnidarian Venoms; D017931:DNA Primers; D018076:DNA, Complementary; D006460:Hemolysin Proteins; D009930:Organic Chemicals; D011994:Recombinant Proteins; C404794:sticholysin II; C121348:stycholysin I