Proteolytic cleavage reveals interaction patterns between silica nanoparticles and two variants of human carbonic anhydrase.
Proteolytic cleavage reveals interaction patterns between silica nanoparticles and two variants of human carbonic anhydrase.
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蛋白水解揭示了二氧化硅纳米粒子和人类碳酸酐酶的两种变体之间的相互作用模式。
DOI：
10.1021/la050477u
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
B. Jonsson
中科院分区：
文献类型：
--
作者：
M. Lundqvist;Cecilia Andresen;S. Christensson;S. Johansson;M. Karlsson;K. Broo;B. Jonsson
为了表征蛋白质表面上参与吸附到二氧化硅纳米颗粒和随后的蛋白质/纳米颗粒相互作用的重排的位点，已经使用了一种新的方法。将蛋白质与二氧化硅纳米颗粒孵育2或16 h后，用胰蛋白酶切割蛋白质，并用质谱法分析肽片段。纳米颗粒表面积超过可用蛋白质表面16倍，以使初始结合受其他蛋白质分子影响的可能性最小化。当在二氧化硅纳米粒子的存在和不存在下获得的片段模式进行比较时，我们能够表征与表面相互作用的蛋白质片段。这种方法使我们能够识别蛋白质结构上的初始结合位点以及与表面长时间孵育后发生的结合位点的重排。
To characterize the sites on the protein surface that are involved in the adsorption to silica nanoparticles and the subsequent rearrangements of the protein/nanoparticle interaction, a novel approach has been used. After incubation of protein with silica nanoparticles for 2 or 16 h, the protein was cleaved with trypsin and the peptide fragments were analyzed with mass spectrometry. The nanoparticle surface area was in 16-fold excess over available protein surface to minimize the probability that the initial binding would be affected by other protein molecules. When the fragment patterns obtained in the presence and absence of silica nanoparticles were compared, we were able to characterize the protein fragments that interact with the surface. This approach has allowed us to identify the initial binding sites on the protein structure and the rearrangement of the binding sites that occur upon prolonged incubation with the surface.