Significance of the N- and C-terminal regions of CAP-1, a cuticle calcification-associated peptide from the exoskeleton of the crayfish, for calcification.
Calcification-associated peptide (CAP)-1 is considered to play an important role in calcification of the exoskeleton of the crayfish, Procambarus clarkii. In this study, in order to clarify the molecular mechanism of calcification, we constructed expression systems of recombinant molecules of CAP-1 and its related peptides in Escherichia coli, and verified the structure-activity relationship of these peptides. The inhibitory assay on calcium carbonate precipitation and the calcium-binding assay showed that the C-terminal acidic region was most important for both activities. The CD spectra of these peptides varied depending on calcium concentration and showed that the N-terminal region is associated with the peptide conformation. These results indicate that the C-terminal part of CAP-1 may concentrate calcium ions for nucleation and/or interact with calcium carbonate precipitate to control the growth and that the N-terminal part contribute to maintaining the peptide conformation.
Inoue
Hirotaka
H
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Ohira
Tsuyoshi
T
Nagasawa
Hiromichi
H
D000595:Amino Acid Sequence; D000818:Animals; D003400:Astacoidea / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D002113:Calcification, Physiologic; D002118:Calcium / Q000378:metabolism; D002119:Calcium Carbonate / Q000378:metabolism; D011232:Chemical Precipitation; D002942:Circular Dichroism; D017931:DNA Primers / Q000235:genetics; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D010455:Peptides; D011994:Recombinant Proteins; C448763:calcification-associated peptide-1, crayfish; D002119:Calcium Carbonate; D002118:Calcium