Biological properties of adrenomedullin conjugated with polyethylene glycol

Kubo, Keishi; Tokashiki, Mariko; Kuwasako, Kenji; Tamura, Masaji; Tsuda, Shugo; Kubo, Shigeru; Yoshizawa‐Kumagaye, Kumiko; Kato, Jun; Kïtamura, Kazuo

doi:10.1016/j.peptides.2014.05.005

Cited by 31 publications

(38 citation statements)

References 21 publications

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“…Albeit there is ongoing improvement in chemical peptide synthesis strategies, it is still a challenge to produce and modify peptides comprising difficult and long peptide sequences like the 52 amino acid ADM with reasonable yields . For the efficient synthesis of the fluorescently labeled [K 9 (Tam)]ADM(1–52) analogue by SPPS, it was necessary to employ resins with low loadings and an additional PEG‐spacer between the Rink amide linker and the resin scaffold to allow the growing peptide chain to move freely during further elongation and modification steps.…”

Section: Discussionmentioning

confidence: 99%

Fluorescently labeled adrenomedullin allows real-time monitoring of adrenomedullin receptor trafficking in living cells

et al. 2015

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The human adrenomedullin (ADM) is a 52 amino acid peptide hormone belonging to the calcitonin family of peptides, which plays a major role in the development and regulation of cardiovascular and lymphatic systems. For potential use in clinical applications, we aimed to investigate the fate of the peptide ligand after binding and activation of the adrenomedullin receptor (AM1), a heterodimer consisting of the calcitonin receptor-like receptor (CLR), a G protein-coupled receptor, associated with the receptor activity-modifying protein 2 (RAMP2). Full length and N-terminally shortened ADM peptides were synthesized using Fmoc/tBu solid phase peptide synthesis and site-specifically labeled with the fluorophore carboxytetramethylrhodamine (Tam) either by amide bond formation or copper(I)-catalyzed azide alkyne cycloaddition. For the first time, Tam-labeled ligands allowed the observation of co-internalization of the whole ligand-receptor complex in living cells co-transfected with fluorescent fusion proteins of CLR and RAMP2. Application of a fluorescent probe to track lysosomal compartments revealed that ADM together with the CLR/RAMP2-complex is routed to the degradative pathway. Moreover, we found that the N-terminus of ADM is not a crucial component of the peptide sequence in terms of AM1 internalization behavior.

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Section: Discussionmentioning

confidence: 99%

Fluorescently labeled adrenomedullin allows real-time monitoring of adrenomedullin receptor trafficking in living cells

et al. 2015

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Section: Molecular Modification Of Ammentioning

confidence: 99%

“…However, AM administered exogenously rapidly disappeared within a few minutes from the blood in animal studies (Ishiyama et al, 1995;Kubo et al, 2014). The first phase of the plasma half-life of AM injected intravenously was 0.62 min in rats (Kubo et al, 2014). In addition, even when infused continuously, the AM dose needs to be monitored carefully because the potent vasodilatory effect would result in inappropriate reduction of blood pressure and activation of the sympathetic nervous system (Kita et al, 2010).…”

Section: Molecular Modification Of Ammentioning

confidence: 99%

“…In an attempt to lessen these unfavorable acute effects, Kubo et al (2014) molecularly modified AM with polyethylene glycol (Fig. 4).…”

Section: Molecular Modification Of Ammentioning

confidence: 99%

“…As shown in Fig. 4, essential for the biological activity are amidated tyrosine at the carboxyl-terminal and the ringed structure formed by a disulfide bond between a pair of cysteines (Eguchi et al, 1994); therefore, conjugation was carried out so as not to impair the pharmacological action by keeping these two sites as molecularly intact as possible (Kubo et al, 2014). As a result, the plasma half-life in vivo became 7-to 8-fold longer without lessening the maximum response of cAMP generation in vitro.…”

Section: Molecular Modification Of Ammentioning

confidence: 99%

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Bench-to-bedside pharmacology of adrenomedullin

Kato

Kïtamura

2015

European Journal of Pharmacology

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Adrenomedullin alleviates mucosal injury in experimental colitis and increases claudin‐4 expression in the colonic epithelium

et al. 2023

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Adrenomedullin (AM) is a peptide with pleiotropic physiological functions that attenuates intestinal mucosal inflammation. However, the mechanism underpinning mucosal protection by AM is not fully understood, and its effect on intestinal epithelial cells remains unclear. Here, we investigated the effects of AM on junctional molecules in primary‐cultured murine intestinal epithelial cells and discovered that AM upregulates claudin‐4 expression. In a mouse model of dextran sulfate sodium‐induced colitis, AM administration also enhanced claudin‐4 expression and accelerated mucosal regeneration. Furthermore, AM reversed TNFα‐mediated downregulation of claudin‐4 and loss of cell–cell adhesion of the HCT116 human intestinal epithelial cell line in vitro. These results indicate that AM may enhance intestinal epithelial integrity by upregulating claudin‐4 expression.

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Biological properties of adrenomedullin conjugated with polyethylene glycol

Cited by 31 publications

References 21 publications

Fluorescently labeled adrenomedullin allows real-time monitoring of adrenomedullin receptor trafficking in living cells

Fluorescently labeled adrenomedullin allows real-time monitoring of adrenomedullin receptor trafficking in living cells

Bench-to-bedside pharmacology of adrenomedullin

Adrenomedullin alleviates mucosal injury in experimental colitis and increases claudin‐4 expression in the colonic epithelium

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