Insight into the collagen assembly in the presence of lysine and glutamic acid: An in vitro study

- The effects of two different charged amino acids in collagen chains on the collagen fibrillogenesis were evaluated.
- The positively charged lysine could improve the sizes or amounts of self-assembled collagen fibrils.
- The width of D-banding of the collagen-lysine co-fibrils increased steadily after introducing lysine.
- The optimal fibrillogenesis was achieved when the concentration of lysine reached to 1 mM.
- The kinetic and thermodynamic collagen self-assembly were both analyzed.

The aim of this study is to evaluate the effects of two different charged amino acids in collagen chains, lysine and glutamic acid, on the fibrillogenesis process of collagen molecules. The turbidity, zeta potential, and fiber diameter analysis suggest that introducing the positively charged lysine into collagen might improve the sizes or amounts of the self-assembled collagen fibrils significantly. Conversely, the negatively charged glutamic acid might restrict the self-assembly of collagen building blocks into a higher order structure. Meanwhile, the optimal fibrillogenesis condition is achieved when the concentration of lysine reaches to 1 mM. Both scanning electron microscopy (SEM) and atomic force microscope (AFM) analysis indicates that compared to pure collagen fibrils, the reconstructed collagen-lysine co-fibrils exhibit a higher degree of inter-fiber entanglements with more straight and longer fibrils. Noted that the specific D-period patterns of the reconstructed collagen fibrils could be clearly discernible and the width of D-banding increases steadily after introducing lysine. Besides, the kinetic and thermodynamic collagen self-assembly analysis confirms that the rate constants of both the first and second assembly phase decrease after introducing lysine, and lysine could promote the process of collagen fibrillogenesis obeying the laws of thermodynamics.

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