Production and functional characterization of a soluble recombinant molt-inhibiting hormone in the mud crab Scylla olivacea

IF 2.2 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology A-Molecular & Integrative Physiology Pub Date : 2025-08-13 DOI:10.1016/j.cbpa.2025.111920

Wanwipa Ittarat , Patchari Yocawibun , Phimsucha Bunphimpapha , Paveena Tapaneeyaworawong , Parichat Chumtong , Sirinart Techa

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Abstract

The molt-inhibiting hormone gene of the mud crab Scylla olivacea (ScoMIH) was successfully cloned and characterized, encoding a 113-amino-acid precursor with a 78-amino-acid mature peptide. Similar to other Brachyuran crabs, ScoMIH is exclusively expressed in the eyestalk ganglia (ES) with slightly higher expression levels during the intermolt stage. To produce a biologically active recombinant ScoMIH, a fusion construct incorporating thioredoxin (Trx) and a His₆ tag was designed, yielding a soluble 27 kDa protein. Functional assays demonstrated that Trx-ScoMIH effectively suppressed the expression of cytochrome P450 monooxygenases in activated Y-organs, leading to reduced ecdysteroid secretion. Furthermore, in vivo administration of Trx-ScoMIH delayed and prolonged the molting process, mimicking the natural molt cycle observed in intact crabs. This study reports the successful production of a soluble, biologically active recombinant ScoMIH, providing a valuable tool for investigating MIH signaling.

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泥蟹Scylla olivacea可溶性重组脱毛激素的制备及功能表征。

成功克隆并鉴定了泥蟹Scylla olivacea （sccomih）的脱毛抑制激素基因，该基因编码一个113个氨基酸的前体和一个78个氨基酸的成熟肽。与其他短爪蟹相似，ScoMIH仅在眼柄神经节（ES）中表达，在换壳期表达水平略高。为了制备具有生物活性的重组ScoMIH，设计了一种包含硫氧还蛋白（Trx）和His₆标签的融合结构，得到可溶性27 kDa蛋白。功能实验表明，trx - scoih有效抑制y脏器细胞色素P450单加氧酶的表达，导致体外类固醇分泌减少。此外，体内给药trx - scoih延迟和延长了蜕皮过程，模拟了在完整螃蟹中观察到的自然蜕皮周期。本研究报道了一种可溶的、具有生物活性的重组ScoMIH的成功生产，为研究MIH信号传导提供了有价值的工具。

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来源期刊

Comparative Biochemistry and Physiology A-Molecular & Integrative Physiology 生物-动物学

CiteScore

5.00

自引率

4.30%

发文量

155

审稿时长

3 months

期刊介绍： Part A: Molecular & Integrative Physiology of Comparative Biochemistry and Physiology. This journal covers molecular, cellular, integrative, and ecological physiology. Topics include bioenergetics, circulation, development, excretion, ion regulation, endocrinology, neurobiology, nutrition, respiration, and thermal biology. Study on regulatory mechanisms at any level of organization such as signal transduction and cellular interaction and control of behavior are also published.