A recombinant molt-inhibiting hormone of the kuruma prawn has a similar secondary structure to a native hormone: determination of disulfide bond arrangement and measurements of circular dichroism spectra.
In crustaceans, molt-inhibiting hormone (MIH) is presumed to regulate molting through suppressing synthesis and/or secretion of ecdysteroids by the Y-organ. Recently, a recombinant MIH of the kuruma prawn Penaeus japonicus was produced in E. coli. To approximate the secondary structure of native and recombinant MIH of P. japonicus containing six cysteine residues, the arrangements of disulfide bridges in both MIHs were determined by characterizing their enzymatic digests, and their circular dichroism spectra were measured. The arrangements of disulfide bonds in both MIHs were determined to be identical, and they were linked between Cys7 and Cys44, Cys24 and Cys40, and Cys27 and Cys53. The circular dichroism spectra of both MIHs were very close, and demonstrated that they were rich in a-helix. a-Helix contents in native and recombinant MIHs were calculated to be 49.3% and 46.0%, respectively. All these results strongly suggested that the recombinant MIH was folded in the same manner as the native MIH.
Katayama
H
H
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Japan.
Ohira
T
T
Nagata
K
K
Nagasawa
H
H
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D033364:Decapoda / Q000737:chemistry*; D004220:Disulfides / Q000737:chemistry; D010450:Endopeptidases / Q000737:chemistry; D006868:Hydrolysis; D007447:Invertebrate Hormones / Q000737:chemistry*; D013058:Mass Spectrometry; D008666:Metalloendopeptidases; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry; D014357:Trypsin / Q000737:chemistry
D004220:Disulfides; D007447:Invertebrate Hormones; D011994:Recombinant Proteins; C012445:molt-inhibiting hormone; D010450:Endopeptidases; D014357:Trypsin; D008666:Metalloendopeptidases; C058921:endoproteinase Asp-N