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钌（II）多吡啶配合物标记的胶原蛋白的物理化学表征研究

Physico-chemical characterization studies of collagen labelled with Ru(II) polypyridyl complex.

作者信息

Anithabanu P, Balasubramanian Surabhya, David Dayanidhi P, Nandhini T, Vaidyanathan V G

机构信息

Advanced Materials Laboratory, CSIR-Central Leather Research Institute, Adyar, Chennai 600020, India.

Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India.

出版信息

Heliyon. 2022 Aug 11;8(8):e10173. doi: 10.1016/j.heliyon.2022.e10173. eCollection 2022 Aug.

DOI:10.1016/j.heliyon.2022.e10173
PMID:36033328
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9404281/
Abstract

The rich luminescence behaviour exerted by transition metal complexes has found significant role in the development of biomolecular and cellular probes. The conjugation of fluorophore to a protein has its own advantage over the label-free system due to its high sensitivity. While numerous proteins have been labelled with either organic or inorganic fluorophores, the conjugation of luminescent transition metal complexes with collagen has not yet been attempted. Here, in this study, the conjugation of a Ru(II) polypyridyl complex with collagen was carried out and its physico-chemical characterization was studied. The conjugation of Ru(II) to collagen was characterized by UV-Visible, fluorescence and ATR-FT-IR spectroscopy. The conjugation of Ru(II) did not alter the triple helical structure of the collagen as evidenced from CD spectral data. The luminescence behaviour of the Ru-tagged collagen was found to be similar to that of the commercially available fluorescein isothiocyanate (FITC) tagged collagen with increase in luminescence upon addition of collagenase. Gel-based collagenase assay showed that the digestion of collagen can be vizualized using UV light due to intrinsic fluorophore tag without carrying out the staining-destaining processes. Energy dispersive X-Ray analysis (EDAX) confirms the presence of Ru in Ru-collagen fibrils. To the best of our knowledge, this is the first report on the conjugation of a Ru(II) complex with the fibrous protein collagen that exhibits similar property as of FITC-collagen and can be used as an alternative.

摘要

过渡金属配合物所展现出的丰富发光行为在生物分子和细胞探针的开发中发挥了重要作用。由于其高灵敏度，荧光团与蛋白质的共轭相对于无标记系统具有自身优势。虽然众多蛋白质已被有机或无机荧光团标记，但发光过渡金属配合物与胶原蛋白的共轭尚未尝试。在此研究中，进行了钌（II）多吡啶配合物与胶原蛋白的共轭，并对其物理化学特性进行了研究。通过紫外可见光谱、荧光光谱和衰减全反射傅里叶变换红外光谱对钌（II）与胶原蛋白的共轭进行了表征。圆二色光谱数据表明，钌（II）的共轭并未改变胶原蛋白的三螺旋结构。发现钌标记的胶原蛋白的发光行为与市售异硫氰酸荧光素（FITC）标记的胶原蛋白相似，添加胶原酶后发光增强。基于凝胶的胶原酶测定表明，由于内在荧光团标记，无需进行染色 - 脱色过程，利用紫外光即可观察到胶原蛋白的消化。能量色散X射线分析（EDAX）证实了钌 - 胶原蛋白原纤维中钌的存在。据我们所知，这是关于钌（II）配合物与纤维状蛋白质胶原蛋白共轭的首次报道，该共轭物表现出与FITC - 胶原蛋白相似的性质，可作为替代品使用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/af5178ec1466/gr10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/40e59e233824/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/f84a99c8a57d/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/e8f4451d5251/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/039506d080fc/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/3244630bdd94/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/f7c0d09540a5/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/0fde857c15c3/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/a923fdcc3e35/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/a104f6038412/gr9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/af5178ec1466/gr10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/40e59e233824/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/f84a99c8a57d/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/e8f4451d5251/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/039506d080fc/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/3244630bdd94/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/f7c0d09540a5/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/0fde857c15c3/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/a923fdcc3e35/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/a104f6038412/gr9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cb/9404281/af5178ec1466/gr10.jpg

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Fluorescent Labeling of Rat-tail Collagen for 3D Fluorescence Imaging.用于三维荧光成像的大鼠尾胶原荧光标记
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