Molecular cloning of the cDNA for the large subunit of the high-Ca2+-requiring form of human Ca2+-activated neutral protease.
A nearly full-length cDNA clone for the large subunit of high-Ca2+-requiring Ca2+-activated neutral protease (mCANP) from human tissues has been isolated. The deduced protein, determined for the first time as an mCANP, has essentially the same structural features as those revealed previously for the large subunits of the low-Ca2+-requiring form (muCANP) [Aoki, K., Imajoh, S., Ohno, S., Emori, Y., Koike, M., Kosaki, G., & Suzuki, K. (1986) FEBS Lett. 205, 313-317] and chicken CANP [Ohno, S., Emori, Y., Imajoh, S., Kawasaki, H., Kisaragi, M., & Suzuki, K. (1984) Nature (London) 312, 566-570]. Namely, the protein, comprising 700 amino acid residues, is characterized by four domains, containing a cysteine protease like domain and a Ca2+-binding domain. The overall amino acid sequence similarities of the mCANP large subunit with those of human muCANP and chicken CANP are 62% and 66%, respectively. These values are slightly lower than that observed between muCANP and chicken CANP (70%). Local sequence similarities vary with the domain, 73-78% in the cysteine protease like domain and 48-65% in the Ca2+-binding domain. These results suggest that CANPs with different Ca2+ sensitivities share a common evolutionary origin and that their regulatory mechanisms are similar except for the Ca2+ concentrations required for activation.
Imajoh
S
S
Department of Molecular Biology, Tokyo Metropolitan Institute of Medical Science, Japan.
Aoki
K
K
Ohno
S
S
Emori
Y
Y
Kawasaki
H
H
Sugihara
H
H
Suzuki
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism; D002154:Calpain / Q000235:genetics* / Q000378:metabolism; D002645:Chickens; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D005796:Genes; D006801:Humans; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009132:Muscles / Q000201:enzymology; D009693:Nucleic Acid Hybridization; D011487:Protein Conformation; D015183:Restriction Mapping; D013154:Spleen / Q000201:enzymology
D046911:Macromolecular Substances; D004247:DNA; D002154:Calpain; D002118:Calcium