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Production of recombinant CART peptides in Escherichia coli with agonist and antagonist effects on food intake in rats

机译：大肠杆菌中重组CART肽的产生对大鼠食物摄取具有激动剂和拮抗剂作用

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摘要

CART (Cocaine- and Amphetamine-Regulated Transcript) peptides modulate food intake and psychostimulant drug actions. Several CART peptides that contain multiple disulfide bonds were produced by overexpression in Escherichia coli bacteria as fusion products with a C-terminal histidine tag. Since these peptides were found denatured in inclusion bodies, in vitro refolding was used to reconstitute their biological activity. These CART peptides were tested for their ability to inhibit food intake in rats. Of recombinant rat CART peptides 1-102 long, 1-102 short, and 55-102, only CART peptide 55-102 dose-dependently inhibited neuropeptide Y stimulated food intake in rats. This effect was dependent on refolding of the peptide since the unfolded version was unable to inhibit food intake. Unfolded CART peptide 55-102 HIS tag, but not CART peptide 1-102 HIS tag, stimulated food intake in rats treated with NPY or vehicle. This effect of unfolded CART peptide 55-102 was consistent with that of an antagonist and is the first description of one. These studies showed that production of CART peptides by bacterial expression is a viable alternative to current methods. (C) 2003 Elsevier Inc. All rights reserved. [References: 18]
机译：CART（可卡因和苯丙胺调节的转录物）肽调节食物摄入和精神刺激药物作用。含有多个二硫键的几种CART肽是通过在大肠杆菌中过表达而产生的，具有C端组氨酸标签的融合产物。由于发现这些肽在包涵体中变性，因此使用体外重折叠来重建其生物学活性。测试了这些CART肽抑制大鼠食物摄入的能力。在重组大鼠CART肽长1-102，短1-102和55-102中，只有CART肽55-102剂量依赖性地抑制了神经肽Y刺激了大鼠的食物摄取。该作用取决于肽的重折叠，因为未折叠的形式不能抑制食物的摄入。未折叠的CART肽55-102 HIS标签，而不是CART肽1-102 HIS标签，刺激了用NPY或赋形剂治疗的大鼠的食物摄入。未折叠的CART肽55-102的这种作用与拮抗剂的作用是一致的，并且是对它的第一个描述。这些研究表明，通过细菌表达产生CART肽是当前方法的可行替代方案。 （C）2003 Elsevier Inc.保留所有权利。 [参考：18]

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