A convenient method for preparation of biologically active recombinant CHH of the kuruma prawn, Marsupenaeus japonicus, using the bacterial expression system

Nagai, Chiaki; Asazuma, Hideaki; Nagata, Shinji; Ohira, Tsuyoshi; Nagasawa, Hiromichi

doi:10.1016/j.peptides.2008.07.017

Cited by 23 publications

(17 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Injection of 300 ng of the purified Mee-CHH-C increased hemolymph glucose levels significantly, and its efficacy was comparable to that of the extract at a concentration of one sinus gland equivalent. In our previous study, injection of 30 pmol (250 ng) of a natural Pej-SGP-VII, one of the six CHHs in M. japonicus, showed almost the same hyperglycemic activity as that of the extract from a single sinus gland by the same assay [23]. These results indicate that Mee-CHH-C has almost the same hyperglycemic activity as that of Pej-SGP-VII in M. japonicus, although Mee-CHH-C is produced in heterogeneous shrimp species M. ensis.…”

Section: Discussionmentioning

confidence: 73%

Isolation and characterization of an additional crustacean hyperglycemic hormone from the greasyback shrimp Metapenaeus ensis

Ohira

Suitoh²,

Yamane

et al. 2010

Fish Sci

Self Cite

View full text Add to dashboard Cite

Crustacean hyperglycemic hormone (CHH) is released from the X-organ/sinus gland complex located in the eyestalks. In this study, the most abundant CHH in the sinus gland of the greasyback shrimp Metapenaeus ensis was purified by reversed-phase HPLC and identified by Nterminal amino acid sequencing. Although two CHH molecules (Mee-CHH-A and Mee-CHH-B) have already been identified from M. ensis by cDNA cloning, this study revealed the presence of an additional CHH peptide based on differences in the N-terminal amino acid sequences of the CHH-A and CHH-B. Therefore, this novel CHH was designated as Mee-CHH-C. A cDNA encoding the Mee-CHH-C precursor was cloned by RT-PCR coupled with 5 0 -and 3 0 -RACE, and it was found that the mature Mee-CHH-C consisted of 72 amino acid residues containing 6 conserved cysteine residues and possessed an amidated C terminus. Mee-CHH-C had 62 and 68% identities with Mee-CHH-A and Mee-CHH-B, respectively, and was highly homologous to CHHs characterized from other penaeid shrimp species. The hyperglycemic activity of Mee-CHH-C was examined by an in vivo bioassay using the kuruma prawn Marsupenaeus japonicus. Injection of Mee-CHH-C increased hemolymph glucose levels significantly and dose-dependently. These results indicate that Mee-CHH-C is possibly one of the major molecules in M. ensis that regulate glucose levels in the hemolymph.

show abstract

Section: Discussionmentioning

confidence: 73%

Isolation and characterization of an additional crustacean hyperglycemic hormone from the greasyback shrimp Metapenaeus ensis

Ohira

Suitoh²,

Yamane

et al. 2010

Fish Sci

Self Cite

View full text Add to dashboard Cite

show abstract

“…This was because, the expressed recombinant Pej-SGP-I aggregated in an insoluble form, consequently yielding an inefficient refolding reaction. In order to overcome this problem, we recently introduced a new system using a Nus-tag for solubilization to produce a recombinant Pej-SGP-VII (Nagai et al 2009). Therefore, soluble and biologically active recombinant Pej-SGP-VII could successfully be obtained using a simpler and more efficient procedure than that previously used for the recombinant Pej-SGP-I.…”

Section: -1e Preparation Of Recombinant Chhsmentioning

confidence: 99%

“…In contrast, the putative receptor of MIH in the Y-organ that was chemically cross-linked with Recently, it was found that the Halloween genes encoding cytochrome P450 monooxygenases from insects were characterized as the steroidogenic enzymes catalyzing the final four steps of the ecdysteroid synthetic pathway from 5β-ketodiol to 20-hydroxyecdysone (Gilbert 2004;Truman 2005). To elucidate whether ecdysteroidogenesis in the Y-organs is regulated by MIH, the expression levels of an orthologue of a member of the Halloween genes, phantom, in the Y-organ of M. japonicus was analyzed (Asazuma et al 2009). As expected, phantom gene expression significantly increased at the pre-molt stage, and decreased after ecdysis (Fig.…”

Section: -2g Mode Of Actionmentioning

confidence: 99%

“…This difference is not yet well understood, and differences in signal transduction systems from hormone reception to biosynthesis of molting hormone should be clarified in the future. Recently, some biosynthetic enzymes involved in the biosynthetic pathway from cholesterol to ecdysone have been characterized at the cDNA level in both insects (Gilbert 2004;Truman 2005;Rewitz and Gilbert 2008) and crustaceans (Rewitz and Gilbert 2008;Asazuma et al 2009).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Crustacean Peptide Hormones: Structure, Gene Expression and Function

Katayama¹,

Ohira²,

Nagasawa³

2013

Aqua-BioSci. Monogr. (ABSM)

Self Cite

View full text Add to dashboard Cite

Keywords AbstractHomeostasis, growth and reproduction in crustaceans are under endocrine control as is established in other animals. The identification of crustacean hormones and elucidation of mechanisms of hormonal regulation have been pursued extensively. Mechanisms are found to be partly similar to, but also considerably different from those in insects, although both groups of animals are arthropods. Molting in insects, for example, is positively regulated by prothoracicotropic hormone (PTTH), which is secreted by the brain/ corpora allata complex and stimulates the prothoracic glands to promote the synthesis and secretion of ecdysteroids. On the other hand, molting in crustaceans is negatively regulated by molt-inhibiting hormone (MIH), which is secreted by the X-organ/sinus gland complex and suppresses the Y-organs from synthesizing ecdysteroids during the intermolt period. Therefore, it is rather difficult to fully elucidate physiological mechanisms in crustaceans based only on the results obtained in insects. Using experimental evidence, we have thus far looked into the roles of crustacean eyestalk hormones and androgenic gland hormone (AGH). Various eyestalk hormones have been isolated from the sinus glands and characterized: e.g., crustacean hyperglycemic hormone (CHH), MIH, vitellogenesis-inhibiting hormone (VIH), red pigment concentrating hormone (RPCH) and pigment dispersing hormone (PDH). CHH, MIH and VIH are structurally similar, forming the CHH-family. Insects also possess molecules similar to CHH, RPCH and PDH, but their functions are different. In addition, AGH has been studied mainly using terrestrial isopods. Here, we describe the present state of our understanding of the structure and function of these peptide hormones, and discuss future perspectives. mercial importance in fisheries and aquaculture, while others play a key role as intermediates in the food chain, linking phytoplanktons and fishes within the aquatic ecosystem. Modern shrimp culture was first established in Japan over 50 years ago using the kuruma prawn Marsupenaeus (formerly Penaeus) japonicus, which is one of the most important commercial species in Japan, and the aquaculture technique has now spread to many countries, especially in tropical and subtropical

show abstract

“…Since the kuruma prawn, M. japonicus, is an important species for aquaculture in Japan, a number of studies on molecular endocrinology such as glucose metabolism, molting regulation, and ovarian maturation have been performed (Yang et al, , 1997Tsutsui et al, 2000;Nagai et al, 2008). In M. japonicus, MIH was purified and characterized as a negative regulator of the ecdysteroidogenesis in the Y-organ .…”

Section: Introductionmentioning

confidence: 99%

Inhibitory effect of molt‐inhibiting hormone on phantom expression in the Y‐organ of the kuruma prawn, Marsupenaeus japonicus

Asazuma

Nagata

Nagasawa

2009

Arch Insect Biochem Physiol

Self Cite

View full text Add to dashboard Cite

Molting in crustaceans is induced by ecdysteroids as in insects. The ecdysteroid titre in hemolymph is negatively regulated by molt-inhibiting hormone (MIH) that inhibits the secretion of ecdysteroids from the Y-organ, an ecdysteroid-producing gland of crustaceans, whereas little is known about the molecular mechanism of inhibition by MIH. Recently, the Halloween genes encoding cytochrome P450 monooxygenases were characterized as the steroidogenic enzymes in insects. To elucidate whether the ecdysteroidogenesis in the Y-organ is regulated by molt-inhibiting hormone (MIH), we analyzed the expression level of an orthologue of a member of the Halloween genes, phantom (Cyp306a1, phm), in the Y-organ of a decapod crustacean, Marsupenaeus japonicus. A cDNA encoding phm (Mj-phm) was cloned by degenerate PCR and 5'- and 3'-RACEs. The deduced amino acid sequence of Mj-phm showed about 40% identity to those of insect phm. The six motif sequences and the four substrate recognition sites were well conserved between Mj-PHM and other PHM. RT-PCR showed the specific expression of Mj-phm mRNA in the Y-organ. In addition, quantitative real-time PCR verified that the expression level of Mj-phm was significantly increased at the pre-molt stage and decreased after ecdysis. Furthermore, exposure of the Y-organ to MIH significantly decreased the Mj-phm expression level in vitro. These results indicate that the transcription of Mj-phm in the Y-organ may be regulated by the inhibitory mechanism of MIH of M. japonicus, which involves the consequent negative regulation of ecdysteroidogenesis at the transcriptional level.

show abstract

A convenient method for preparation of biologically active recombinant CHH of the kuruma prawn, Marsupenaeus japonicus, using the bacterial expression system

Cited by 23 publications

References 42 publications

Isolation and characterization of an additional crustacean hyperglycemic hormone from the greasyback shrimp Metapenaeus ensis

Isolation and characterization of an additional crustacean hyperglycemic hormone from the greasyback shrimp Metapenaeus ensis

Crustacean Peptide Hormones: Structure, Gene Expression and Function

Inhibitory effect of molt‐inhibiting hormone on phantom expression in the Y‐organ of the kuruma prawn, Marsupenaeus japonicus

Contact Info

Product

Resources

About