Hisactophilin, a histidine-rich actin-binding protein from Dictyostelium discoideum.
The purification, cloning, and complete cDNA-derived sequence of a 17-kDa protein of Dictyostelium discoideum are described. This protein binds to F-actin in a pH-dependent and saturable manner. It induces actin polymerization in the absence of Mg2+ or K+, and is enriched in the submembranous region of the amoeboid cells as indicated by immunofluorescence labeling of cryosections. The mRNA as well as the protein are present throughout growth and all stages of development. The protein is detected in both soluble and particulate fractions of the cells. From a plasma membrane-enriched fraction, minor amounts of the protein are stepwise solubilized with 1.5 M KCl, 0.1 M NaOH, and Triton X-100, but most of the protein is only solubilized with 1% sodium dodecyl sulfate. As judged by the apparent molecular mass in sodium dodecyl sulfate-polyacrylamide gels, immunological cross-reactivity, and two-dimensional electrophoresis, the 17-kDa proteins from the soluble and particulate fraction resemble each other. The cDNA sequence does not reveal any signal peptide, trans-membrane region, or N-glycosylation site. Southern blots hybridized with a cDNA probe that spans the entire coding region show that the 17-kDa protein is encoded by a single gene. The most characteristic feature of the protein is its high content of 31 histidine residues out of 118 amino acids. We designate this protein as hisactophilin and suggest that this histidine-rich protein responds in its actin-binding activity to changes in cellular pH upon chemotactic signal reception.
Scheel
J
J
Max-Planck-Institute für Biochemie, Martinsried, Federal Republic of Germany.
Ziegelbauer
K
K
Kupke
T
T
Humbel
B M
BM
Noegel
A A
AA
Gerisch
G
G
Schleicher
M
M
D000199:Actins / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics; D004023:Dictyostelium / Q000235:genetics / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D005796:Genes; D005800:Genes, Fungal; D046911:Macromolecular Substances; D008840:Microfilament Proteins; D008969:Molecular Sequence Data; D008970:Molecular Weight; D015800:Protozoan Proteins; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic
D000199:Actins; D002352:Carrier Proteins; D004271:DNA, Fungal; D005656:Fungal Proteins; D046911:Macromolecular Substances; D008840:Microfilament Proteins; D015800:Protozoan Proteins; D012333:RNA, Messenger; C058561:hisactophilin protein, Protozoan