A convenient method for preparation of biologically active recombinant CHH of the kuruma prawn, Marsupenaeus japonicus, using the bacterial expression system.
Crustacean hyperglycemic hormone (CHH) not only plays an important role in the modulation of hemolymph glucose level but also functions in other biological events including molting, reproduction and stress response. Of the six CHHs characterized in Marsupenaeus japonicus, an expression system for recombinant Pej-SGP-VII (rPej-SGP-VII-amide) has not yet been established. Here, we established a procedure using a Nus-tag for solubilization, thereby soluble and biologically active rPej-SGP-VII-amide could successfully be obtained by a simpler procedure than previous ones used for producing other recombinant Pej-SGPs (Pej-SGP-I, III and IV). It was found that rPej-SGP-VII-amide thus obtained had the correct arrangement of intramolecular disulfide bonds and helix-rich secondary structure. The established expression system for rPej-SGP-VII-amide may be applicable for the preparation of other recombinant CHHs.
Nagai
Chiaki
C
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.
Asazuma
Hideaki
H
Nagata
Shinji
S
Ohira
Tsuyoshi
T
Nagasawa
Hiromichi
H
D000595:Amino Acid Sequence; D000818:Animals; D060829:Arthropod Proteins; D001681:Biological Assay; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D006943:Hyperglycemia / Q000139:chemically induced; D007447:Invertebrate Hormones; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000302:isolation & purification*; D033561:Penaeidae / Q000737:chemistry*; D011994:Recombinant Proteins / Q000302:isolation & purification
D060829:Arthropod Proteins; D007447:Invertebrate Hormones; D009419:Nerve Tissue Proteins; D011994:Recombinant Proteins; C015947:hyperglycemic hormone, crustacean