Evidence for structural differences between the two highly homologous actin-regulatory proteins, destrin and cofilin.
The amino acid sequences of destrin and cofilin are very similar (84% homology) throughout the entire range of proteins, but they have different functions. In this study, we constructed a new cofilin expression plasmid, which had high expression frequency, and the structures of destrin and cofilin were analyzed by limited proteolysis and circular dichroism (CD). When destrin was digested by trypsin, two fragments of 17.0 kDa and 9.2 kDa were obtained, whereas only one 8.4 kDa fragment was obtained from cofilin. In spite of the overall sequence homology, an N-terminal amino acid sequence analyses of the fragments revealed the cleavage sites on destrin and cofilin to be different. These results suggest that destrin and cofilin differ in their overall tertiary folds. Cofilin showed activity similar to destrin at high pH values, although no pH-dependent structural change in cofilin was confirmed by using limited proteolysis and CD.
Arima
K
K
Department of Biochemical Engineering and Science, Faculty of Computer Science and Systems Engineering, Kyushu Institute of Technology, Fukuoka, Japan. arima@earth83.bse.kyutech.ac.jp
Imanaka
M
M
Okuzono
S
S
Kazuta
Y
Y
Kotani
S
S
D051339:Actin Depolymerizing Factors; D000199:Actins / Q000737:chemistry*; D000595:Amino Acid Sequence; D002352:Carrier Proteins / Q000737:chemistry*; D002918:Chymotrypsin / Q000737:chemistry; D002942:Circular Dichroism; D003598:Cytoskeletal Proteins; D051345:Destrin; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D008840:Microfilament Proteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000737:chemistry; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D013381:Subtilisins / Q000737:chemistry; D014357:Trypsin / Q000737:chemistry
D051339:Actin Depolymerizing Factors; D000199:Actins; D002352:Carrier Proteins; D003598:Cytoskeletal Proteins; D051345:Destrin; D008840:Microfilament Proteins; D013381:Subtilisins; D002918:Chymotrypsin; D014357:Trypsin