Nanoscience and Nanotechnology Letters
·
2019
·Vol. 11
·期 8
·1104-1115
·
IF 0
Bhatti Huma,Jawed Rohil,Liu Hang,Ali Irshad,Zhu Libo,Liu Quanjun
Biological nanopores have been proved as impressive nano-devices capable of holding the detection sensitivity from a nanoparticle to a single macromolecule. Protein pores are efficiently reproducible with assured channel geometry and their propensity to be engineered easily make them
convenient in nanopore sensing techniques. Among various porin-proteins, MspA porin of Mycobacterium smegmatis is quite ideal for nanopore DNA sequencing. However, the traversing speed of singlestranded DNA (ssDNA) through both the mutants of MspA (M1MspA and M2MspA) is too fast to
be analyzed. Therefore, we utilized a new mutant of MspA, termed as M3MspA which was previously purified in our laboratory. We, for the first time, demonstrated that translocation of ssDNA is achieved at a minimum voltage-threshold of 40 mV at pH 8.0. This new mutant of MspA exhibits a crystal-clear
discrimination between two homopolymers, poly dA50 and poly dC50 with duration time up to few milliseconds and well-resolved signature current-amplitudes were obtained for both sides (3′ or 5′) of the polymer entered the pore, without any modification to ssDNA.
Significantly, M3MspA determines a precise length of a hetero-homopolymer, poly(dA50dC50) by producing a dwell time congruent to the sum of the time spent by their component-homopolymers, separately. On account of their specificity and sensitivity towards translocating
polynucleotides, we suggest that this new mutant of MspA would be a sophisticated bio-nanopore in further enlightening the translocation dynamics caused by diverse conformations of a same biomolecule. On top of that, it would also be helpful in nanopore DNA sequencing.
研究领域:
生物物理学
·
纳米技术
·
分子生物学
·
基因组学
✓ 已验证
DOI
Matter
·
2024
·
IF 17.3
Pingping Fan,Kui Li,Tian Li,Panke Zhang,Shuo Huang
Alcoholic beverages, such as wine, beer, and distilled spirits, are widely produced and consumed in different nations. Different types of alcoholic beverages contain different combinations of flavor compounds. However, rapid and simultaneous analysis of a large variety of compounds in alcoholic beverages by a miniatured and portable device remains a challenge. In this paper, a Mycobacterium smegmatis porin A (MspA) nanopore modified with a phenylboronic acid (PBA) adapter is applied for rapid analysis of a variety of alcoholic beverages. By utilizing custom machine learning algorithms, various cis -diols are identified simultaneously in both distilled and fermented alcoholic beverages, generating unique barcodes for each sample type. Nanopore analysis of alcoholic beverages has also never been carried out previously. Rapid grading of wine sweetness and detection of additives, including sucrose and D-tartaric acid, are also demonstrated, showcasing the significance of this technique in the administration of wine production.
研究领域:
生物技术
·
食品科学
✓ 已验证
产品关联
DOI
高影响因子
ACS Sensors
·
2021
·
IF 7.711
Jinyue Zhang,Jiao Cao,Wendong Jia,Shanyu Zhang,Shuanghong Yan,Yuqin Wang,Panke Zhang,Hong-Yuan Chen,Wenfei Li,Shuo Huang
Protein nanopores can be engineered as nanoreactors to investigate single-molecule chemical reactions. Recent studies have demonstrated that Mycobacterium smegmatis porin A (MspA) nanopore is a superior engineering template acknowledging its geometrical advantages. However, reported engineering of MspA to form a nanoreactor has focused only on site 91 and mapping of other engineering sites have never been performed before. By taking tetrachloraurate(III) ([AuCl4]−) as a model reactant, potential engineering sites within the pore constriction of MspA have been thoroughly investigated. It is discovered that the produced event amplitude is inversely correlated to the cross-sectional diameter of the pore constriction size at the engineering site, providing evidence that site 91 is actually already the optimum place to introduce the chemical reactivity. Other unavailable engineering sites, which either significantly interfere with the pore assembly or produce reactive sites facing to the pore’s exterior instead of to the pore lumen, were also spotted and discussed. All results demonstrated above have provided a complete map of engineering sites within the constriction area of MspA and may be beneficial as a reference in future engineering of corresponding nanoreactors.
研究领域:
纳米技术
·
生物化学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2020
·
IF 6.785
Jiao Cao,Shanyu Zhang,Jinyue Zhang,Sha Wang,Wendong Jia,Shuanghong Yan,Yuqin Wang,Panke Zhang,Hong-Yuan Chen,Shuo Huang
Gold(I) compounds are known to bind sulfur-containing proteins, forming the basis in the design of gold(I)-based drugs. However, the intrinsic molecular mechanism of the chemical reaction is easily hidden when monitored in ensemble. We have previously demonstrated that Mycobacterium smegmatis porin A (MspA) can be engineered (MspA-M) to contain a specialized nanoreactor to probe chemical reactions involving tetrachloroaurate(III). Here, we provide further investigations of coordination interactions between dichloroaurate(I) and MspA-M. Gold compounds of different coordination geometry and valence states are as well probed and evaluated, demonstrating the generality of MspA-M. With single-molecule evidence, MspA-M demonstrates a preference for dichloroaurate(I) than tetrachloroaurate(III), an observation in a single molecule that has never been reported. By counting the maximum number of simultaneous ion bindings, the narrowly confined pore restriction also efficiently distinguishes dichloroaurate(I) and tetrachloroaurate(III) according to their differences in geometry or size. The above demonstration complemented a previous study by demonstrating other possible gold-based single-molecule chemical reactions observable by MspA. These observations bring insights in the understanding of gold-based coordination chemistry in a nanoscale.
研究领域:
化学
·
纳米技术
·
生物物理学
·
材料科学
✓ 已验证
DOI
PubMed
高影响因子
Matter
·
2025
·
IF 17.5
Kui Li,Tian Li,Kefan Wang,Yifan Wang,Zixuan Wang,Bingxiao Cheng,Yusheng Ouyang,Panke Zhang,Shuo Huang
Urine, an easily accessible body fluid, contains rich metabolic compounds reflecting the human health condition. Though urine analysis can be carried out by many conventional analytical methods, direct, rapid, and simultaneous identification of a variety of urinary metabolites by a handheld device remains challenging. Recent developments in nanopore small molecule sensing may suggest direct urine analysis performed by an engineered nanopore. Here, a hetero-octameric Mycobacterium smegmatis porin A (MspA) nanopore immobilized with a single nickel ion is confirmed to be suitable for the simultaneous identification of six urinary metabolites: creatinine, taurine, histidine, 1-methylhistidine, 3-methylhistidine, and hypoxanthine. With the assistance of machine learning, an overall accuracy of 98.5% was reported. This urine sensing capacity was further applied in real-time urinary metabolite monitoring after diet and physical exercise. These proofs of concept suggest a variety of nanopore-based body fluid analyses for early diagnosis, health management, pharmacokinetic monitoring, and doping tests.
研究领域:
分析化学
·
纳米技术
·
代谢组学
·
生物医学工程
·
机器学习
·
运动科学
·
营养科学
✓ 已验证
产品关联
DOI
高影响因子
ACS Nano
·
2023
·
IF 17.1
Xiaoyu Du,Shanyu Zhang,Liying Wang,Yuqin Wang,Pingping Fan,Wendong Jia,Panke Zhang,Shuo Huang
Isomers of some chemical compounds may be dynamically interconvertible. Due to a lack of sensing methods with a sufficient resolution, however, direct monitoring of such processes can be difficult. Engineered Mycobacterium smegmatis porin A (MspA) nanopores can be applied as nanoreactors so that chemical reactions can be directly monitored. Here, an MspA modified with a phenylboronic acid (PBA) adapter was prepared and was used to observe dynamic interconversion between chiral configurations of boronate esters, which appears as telegraphic switching on top of nanopore events. The mechanism of this behavior was further confirmed by trials with different halogenated catechols, dopamine, adenosine, 1,2-propanediol, and (2R,3R)-2,3-butanediol, and its generality has been demonstrated. These results suggest that an engineered MspA possesses an exceptional resolution in its monitoring of chemical reaction processes and may inspire the future design of nanopore small-molecule sensors.
研究领域:
纳米技术
·
化学生物学
·
分析化学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
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