Cloning and characterization of actin depolymerizing factor from Toxoplasma gondii.
We determined the predicted amino acid sequence of actin depolymerizing factor (ADF) from Toxoplasma gondii by sequencing the full-length cDNA. T. gondii ADF consists of 118 amino acids (calculated molecular weight 13,400) and shares a high degree of sequence similarity to other low molecular weight actin monomer sequestering proteins, especially Acanthamoeba actophorin, plant ADFs and yeast and vertebrate cofilin. ADF from T. gondii is smaller and does not contain a nuclear localization sequence like the related vertebrate proteins. Southern blot analysis indicates that T. gondii ADF is a single-copy gene. Homogeneous recombinant T. gondii ADF purified from E. coli is active in binding actin monomers and depolymerizing F-actin. Localization of ADF by immunofluorescence and immunoelectron microscopy indicates ADF is scattered throughout the cytoplasm and prominently localized beneath the plasma membrane in T. gondii.
Allen
M L
ML
Department of Molecular Pharmacology, Stanford University School of Medicine, CA 94305-5332, USA.
Dobrowolski
J M
JM
Muller
H
H
Sibley
L D
LD
Mansour
T E
TE
D051339:Actin Depolymerizing Factors; D000199:Actins / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics; D051345:Destrin; D017125:Genes, Protozoan; D007150:Immunohistochemistry; D008840:Microfilament Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014122:Toxoplasma / Q000235:genetics* / Q000378:metabolism*
D051339:Actin Depolymerizing Factors; D000199:Actins; D017931:DNA Primers; D018076:DNA, Complementary; D016054:DNA, Protozoan; D051345:Destrin; D008840:Microfilament Proteins; D015800:Protozoan Proteins; D011994:Recombinant Proteins