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Polypeptide analysis for nanopore-based protein identification

Nano Research 2022年第11期15卷 9831-9842页

作者： mazdak afshar bakshloo Safia Yahiaoui Fabien Piguet Manuela Pastoriza-Gallego Régis Daniel Jérôme Mathé John J.Kasianowicz Abdelghani Oukhaled CY Cergy Paris Université CNRSLAMBE95000CergyFrance UniversitéParis-Saclay Univ EvryCNRSLAMBE91025Evry-CourcouronnesFrance University of South Florida Dept.of PhysicsTampaFL 33620USA Freiburg Institute for Advanced Studies Universität FreiburgFreiburg 79104Germany

Presently,proteins are identified by cleaving them with proteases,measuring the mass to charge ratio of the fragments with a mass spectrometer,and matching the fragments to segments within known proteins in *** earlier demonstrated that a nanometer-scale pore formed by aerolysin(AeL)can discriminate between,and therefore identify,three similar size proteins from their trypsin-cleaved polypeptide *** this nanopore-protease method,the protein’s identity is instead determined from characteristic ionic current blockade patterns caused by the polypeptide fragments that enter the *** results also suggested that not all of the theoretically expected cleavage products partition into the *** better understand the mechanism by which polypeptide fragments are captured,and how different polypeptides reduce the pore’s ionic current,we studied the effects of 11 identical length polypeptides with different net charges and charge *** show that under certain experimental conditions,negative,positive,and neutral polypeptides are driven into the AeL pore by the same applied voltage *** capture rate and dwell time of polypeptides in the pore depend strongly on the ionic strength,the magnitude of the applied voltage,and the net charge and charge distribution of the *** dwell time distribution depends nonmonotonically on the applied voltage(regardless of the polymer’s net charge),and its maximum value depends on the polypeptide net charge and charge *** maximum dwell time for different polypeptides does not occur at the same applied voltage amplitude,which conceivably might complicate the detection and discrimination of some polypeptide *** additional experiments,computer simulations,and artificial intelligence research are needed to better understand how to optimize the partitioning of enzymatically cleaved fragments into the AeL nanopore,the method is still capable of accurately identifying

关键词： protein identification nanopores trypsin enzymatic degradation

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