分类： 生物学 >> 生物工程 提交时间： 2024-03-05
Zhang, Ming
Tang, Chao
Wang, Zichun
Chen, Shanchuan
Xu, Mengying
Li, Kaiju
Sun, Ke
Zhao, Changjian
Wang, Yu
Dai, Lunzhi
Lu, Guangwen
Shi, Hubing
Chen, Lu
Geng, Jia
摘要： Single-molecule discrimination among amino acids is crucial to the realization of next-generation protein sequencing. Owing to the heterogeneous charge and subtle volume difference of underivatized amino acids, it remains a challenge for single-molecule techniques to recognize each of them. Here, we report the direct detection of twenty proteinogenic amino acids using a copper(II)-functionalized MspA nanopore. The binding sites for copper(II) ion are constructed by introducing histidine mutation (N91H) to M2MspA protein. With copper ion binding to histidine residues, amino acids can reversibly coordinate the copper-histidine complex, generating well-defined current signals. Using this strategy, all twenty amino acids can be detected. Assisted by a machine learning algorithm, we can identify 100% of signals with 70.2% accuracy or 60% of signals with 93.4% accuracy in the validation set. In successively addition experiment, each amino acid in a mixture of 10 amino acids can be identified precisely. Furthermore, we use carboxypeptidase A1 to partly release the C-terminal amino acids of peptides with different lengths (9, 10 and 22 residues). The hydrolysates of peptides can be identified and distinguished. These results demonstrate the feasibility of this system for amino acids detection and peptide identification, shedding new lights on the development of single-molecule protein sequencing.
分类： 地球科学 >> 空间物理学 提交时间： 2016-05-12
摘要： Based on a hybrid galactic cosmic-ray transport model, which incorporated MHD global heliospheric data into Parker's cosmic-ray transport equation, we studied the behavior of the transport of galactic cosmic rays and the corresponding gradients in their flux near the heliopause (HP). We found that, (1) by increasing the ratio of the parallel diffusion coefficient to the perpendicular diffusion coefficient in the interstellar magnetic field of the outer heliosheath, the simulated radial flux near the HP increases as well. As the ratio multiplying factor reached 1010, the radial flux experienced a sudden jump near the HP, similar to what Voyager 1 observed in 2012. (2) The effect of changing the diffusion coefficients' ratio on the radial flux variation depends on the energy of the cosmic rays, the lower the energy, the more pronounced the effect is. (3) The magnitude of the diffusion coefficients also affect the radial flux near the HP, the modulation beyond the HP varies by adjusting the magnitude multiplying factor.
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