Interaction of Lysozyme with Poly(L-lysine)/Hyaluronic Acid Multilayers: An ATR-FTIR Study.
Interaction of Lysozyme with Poly(L-lysine)/Hyaluronic Acid Multilayers: An ATR-FTIR Study.
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DOI：
10.3390/polym15041036
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发表时间：
2023-02-19
期刊：
影响因子：
5
通讯作者：
Volodkin D
中科院分区：
文献类型：
--
作者：
Velk N;Keller J;Duschl C;Brezesinski G;Volodkin D
载有生物活性分子如蛋白质的聚电解质多层膜（PEM）作为细胞外基质的优异模拟物，并且可以在诸如生物医学和细胞生物学的领域中找到应用。一个问题，这是至关重要的成功就业的质子交换膜的是是否构象和生物活性的负载蛋白质被保存。采用偏振衰减全反射傅里叶变换红外光谱（ATR-FTIR）技术研究了蛋白质溶菌酶（Lys）在聚赖氨酸/透明质酸（PLL/HA）多层膜中的构象。在蛋白质吸附和解吸期间，从涂覆在ATR晶体上的PEM中的蛋白质获取光谱。为了比较，对与未涂覆晶体接触的赖氨酸的情况进行类似的研究。该研究强调了PEM中存在“紧密结合”和“结合不良”的Lys组分。这些级分在它们的构象和在洗涤时从PEM的释放行为上不同。用不同的偏振记录的光谱的比较表明优先取向的α螺旋结构，β片和转向的“紧密结合”的赖氨酸。相反，“结合不良”的部分显示各向同性取向，其构象被很好地保存。
Polyelectrolyte multilayers (PEM) loaded with bioactive molecules such as proteins serve as excellent mimics of an extracellular matrix and may find applications in fields such as biomedicine and cell biology. A question which is crucial to the successful employment of PEMs is whether conformation and bioactivity of the loaded proteins is preserved. In this work, the polarized attenuated total reflection Fourier transform infrared (ATR-FTIR) technique is applied to investigate the conformation of the protein lysozyme (Lys) loaded into the poly(L-lysine)/hyaluronic acid (PLL/HA) multilayers. Spectra are taken from the protein in the PEMs coated onto an ATR crystal during protein adsorption and desorption. For comparison, a similar investigation is performed for the case of Lys in contact with the uncoated crystal. The study highlights the presence of both “tightly” and “poorly bound” Lys fractions in the PEM. These fractions differ in their conformation and release behavior from the PEM upon washing. Comparison of spectra recorded with different polarizations suggests preferential orientation of alpha helical structures, beta sheets and turns in the “tightly bound” Lys. In contrast, the “poorly bound” fraction shows isotropic orientation and its conformation is well preserved.
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