埃德曼退化
体内
肽
青蛙皮
生物
化学
抗菌肽
分馏
高效液相色谱法
两栖动物
生物化学
色谱法
药理学
肽序列
生态学
有机化学
钠
生物技术
基因
作者
J. Michael Conlon,R. Charlotte Moffett,Peter R. Flatt,Jérôme Leprince
出处
期刊:Methods in molecular biology
日期:2024-01-01
卷期号:: 291-306
标识
DOI:10.1007/978-1-0716-3646-6_15
摘要
Several amphibian peptides that were first identified on the basis of their antimicrobial or cytotoxic properties have subsequently shown potential for development into agents for the treatment of patients with Type 2 diabetes. A strategy is presented for the isolation and characterization of such peptides that are present in norepinephrine-stimulated skin secretions from a range of frog species. The methodology involves (1) fractionation of the secretions by reversed-phase HPLC, (2) identification of fractions containing components that stimulate the rate of release of insulin from BRIN-BD11 clonal β-cells without simultaneously stimulating the release of lactate dehydrogenase, (3) identification of active peptides in the fractions in the mass range 1–6 kDa by MALDI-ToF mass spectrometry, (4) purification of the peptides to near homogeneity by further reversed-phase HPLC on various column matrices, and (5) structural characterization by automated Edman degradation. The effect of synthetic replicates of the active peptides on glucose homeostasis in vivo may be evaluated in appropriate animal models of Type 2 diabetes such as db/db mice and mice fed a high fat diet to produce obesity, glucose intolerance, and insulin resistance.
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