Expression of a Recombinant Molt-Inhibiting Hormone of the Kuruma Prawn<i>Penaeus japonicus</i>in<i>Escherichia coli</i>

Ohira, Tsuyoshi; Nishimura, Takayuki; Sonobe, Haruyuki; Okuno, Akira; Watanabe, Toshiki; Nagasawa, Hiromichi; Kawazoe, Ichiro; Aida, Katsumi

doi:10.1271/bbb.63.1576

Cited by 41 publications

(23 citation statements)

References 20 publications

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“…Sample Preparation-A recombinant M. japonicus MIH (r-MIH) was overproduced using a pET-28a(ϩ) plasmid (Novagen) in Escherichia coli BL21-Codonplus™(DE3)-RIL (Stratagene) and purified after the refolding reaction as described previously (5,8). Uniformly Circular Dichroism Spectral Analysis-The refolded r-MIH was dissolved in 30% acetonitrile aqueous solution at the concentration of 100 g/ml.…”

Section: Methodsmentioning

confidence: 99%

The Solution Structure of Molt-inhibiting Hormone from the Kuruma Prawn Marsupenaeus japonicus

Katayama¹,

Nagata²,

Ohira³

et al. 2003

Journal of Biological Chemistry

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Molting in crustaceans is controlled by molt-inhibiting hormone (MIH) and ecdysteroids. It is presumed that MIH inhibits the synthesis and the secretion of ecdysteroids by the Y-organ, resulting in molt suppression. The amino acid sequence of MIH is similar to that of crustacean hyperglycemic hormone (CHH), and therefore, they form a peptide family referred to as the CHH family. Most of the CHH family peptides show no cross-activity, whereas a few peptides show multiple hormonal activities. To reveal the structural basis of this functional specificity, we determined the solution structure of MIH from the Kuruma prawn Marsupenaeus japonicus and compared the solution structure of MIH with a homology-modeled structure of M. japonicus CHH. The solution structure of MIH consisted of five ␣-helices and no ␤-structures, constituting a novel structural motif. The homology-modeled structure of M. japonicus CHH was very similar to the solution structure of MIH with the exception of the absence of the N-terminal ␣-helix and the C-terminal tail, which were sterically close to each other. The surface properties of MIH around this region were quite different from those of CHH. These results strongly suggest that this region is a functionally important site for conferring molt-inhibiting activity.Molting is one of the most significant processes occurring during the arthropod life cycle and is triggered by a molting hormone, ecdysteroids. In crustaceans, it is presumed that the synthesis and the secretion of ecdysteroids by the Y-organ are suppressed by molt-inhibiting hormone (MIH) 1 (1). MIH is produced by the X-organ and released from the sinus gland located in the eyestalk. Crustacean hyperglycemic hormone (CHH), gonad-inhibiting hormone, and mandibular organ-inhibiting hormone are also synthesized in and released from the X-organ/sinus gland complex. Most of these peptides consist of 72-78 amino acid residues and exhibit similar amino acid sequences. Therefore, these peptides form a peptide family referred to as the CHH family (2). The CHH family peptides commonly have six cysteine residues, which form three intramolecular disulfide bonds. Circular dichroism spectral analyses of some CHH family peptides demonstrated that they were rich in ␣-helices (3-5). However, there has so far been no report on the determination of the tertiary structure of any of the CHH family peptides.CHH family peptides show various biological activities: some peptides that suppress molting, increase hemolymph glucose levels, and suppress vitellogenesis in the ovary and others that suppress the synthesis of methyl farnesoate in the mandibular organ (2). In general, each CHH family peptide shows only one type of biological activity, although a few peptides show multiple hormonal activities. For instance, CHH from the American lobster Homarus americanus also exhibits molt-inhibiting activity (6), and mandibular organ-inhibiting hormone from the spider crab Libinia emarginata exhibits hyperglycemic activity (7).To reveal the structural basis of the fu...

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Section: Methodsmentioning

confidence: 99%

The Solution Structure of Molt-inhibiting Hormone from the Kuruma Prawn Marsupenaeus japonicus

Katayama¹,

Nagata²,

Ohira³

et al. 2003

Journal of Biological Chemistry

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“…The construction of expression plasmids was followed as described by Ohira and colleagues (Ohira et al, 1999). In brief, the forward primer (5Ј-ATCCATGGCCCAGATTTACGACTCCTC -CTGT-3Ј, Invitrogen, Carlsbad, CA, USA) contained 14 nucleotide residues encoding QIYDS (the first five amino acid residues of mature PO-CHH).…”

Section: Construction Of Expression Plasmidsmentioning

confidence: 99%

“…The procedure for the expression of rPO-CHH in E. coli Rosettagami(DE3)pLysS competent cells (Novagen) was as described (Ohira et al, 1999). The supernatant and the re-suspended insoluble material were separated by 18% SDS-PAGE, and the rPO-CHH was found in both fractions (supplementary material Fig.…”

Section: Expression Of Recombinant Po-chhmentioning

confidence: 99%

The specific binding sites of eyestalk- and pericardial organ-crustacean hyperglycaemic hormones (CHHs) in multiple tissues of the blue crab,Callinectes sapidus

Katayama

Chung

2009

Journal of Experimental Biology

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SUMMARYCrustacean hyperglycaemic hormone from the pericardial organ (PO-CHH) is a CHH-related neuropeptide but its function and target tissues are not known in crustaceans. To investigate this issue, we employed radiolabelled ligand binding and cGMP assays, using eyestalk-CHH (ES-CHH) as a reference neuropeptide. The membranes were prepared from various tissues of Callinectes sapidus: hepatopancreas, hindgut, midgut, gills, heart, abdominal muscles and scaphognathites. Like ES-CHH, recombinant PO-CHH (rPO-CHH) specifically bound to the membranes of scaphognathites=abdominal muscles>midgut>gills> heart>hindgut and hepatopancreas (list order corresponds to the number of binding sites). The specific binding sites of 125 I-ES-CHH in hepatopancreas and gills were saturable and displaceable. The abdominal muscle membrane binding sites were specific and saturable to both CHHs. These binding sites were displaced by homologous neuropeptides, but poorly displaced by the heterologous counterpart. As for the second messenger, the expected increment (3-to >20-fold) in the amount of cGMP produced by ES-CHH was noted in most tissues tested except midgut. Recombinant PO-CHH increased cGMP production 1.5-to 4-fold in scaphognathites, heart, midgut, hindgut and abdominal muscles. The results obtained from the binding study suggest that PO-CHH also has multiple target tissues of which abdominal muscles and scaphognathites are the primary ones. The differences in the primary amino acid sequences of PO-CHH and ES-CHH, particularly in the C-terminal region and in the amidation at Cterminus, may contribute to the truncated responses of hyperglycaemia, cGMP stimulation and binding affinity. Supplementary material available online at

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“…These assay methods are very sensitive, and will be used for bioassay of the characterization process. After characterization, the factors may be synthesized by molecular biological techniques as recombinant molt-inhibiting hormone and recombinant androgenic gland hormone produced by Ohira et al 13 and Okuno et al 22 . For male reproduction, administration of the androgenic gland will become available, although purification of the hormone in shrimp is required.…”

Section: Perspectives On Hormonal Manipulationmentioning

confidence: 99%

Perspectives on Hormonal Manipulation of Shrimp Reproduction

Okumura

2004

JARQ

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Expression of a Recombinant Molt-Inhibiting Hormone of the Kuruma PrawnPenaeus japonicusinEscherichia coli

Cited by 41 publications

References 20 publications

The Solution Structure of Molt-inhibiting Hormone from the Kuruma Prawn Marsupenaeus japonicus

The Solution Structure of Molt-inhibiting Hormone from the Kuruma Prawn Marsupenaeus japonicus

The specific binding sites of eyestalk- and pericardial organ-crustacean hyperglycaemic hormones (CHHs) in multiple tissues of the blue crab,Callinectes sapidus

Perspectives on Hormonal Manipulation of Shrimp Reproduction

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