Voltage-driven Structural Modulation of Single Enzyme Molecules with Nanopipettes

Abstract

Electric-field modulation of protein structure has attracted increasing interest, yet quantitative characterization under nanoconfinement remains challenging. Here, we investigate the voltage-dependent translocation of formate dehydrogenase (FDH) through a nanopipette. The ionic current blockade increases linearly between 300 and 500 mV before deviating from the low-voltage trend at higher voltages, accompanied by corresponding changes in dwell time and event frequency. Quantitative analysis of ionic current blockades reveals a progressive reduction in the effective excluded volume of FDH, decreasing by approximately 13% at 600 mV and 23% at 700 mV relative to the low-voltage range. These findings suggest the electric field can serve as a modulator of FDH conformation during translocation, highlighting its dual role in both driving transport and modulating protein structure at the single-molecule level.

Keywords

Nanopipette / Single-molecule sensing / Structural modulation

References

[1] Corbella M, Pinto G P, Kamerlin S C LZNat. Rev. Chem., 2023, 7536.

[2] Torgeson K R, Clarkson M W, Granata D, Lindorff-Larsen K, Page R, Peti WZSci. Adv., 2022, 8eabo5546.

[3] Hegazy R, Richard J PZChem. Rev., 2025, 12511524.

[4] Mondal D, Snodgrass H M, Gomez C A, Lewis J CZChem. Rev., 2023, 12310381.

[5] Secundo FZChem. Soc. Rev., 2013, 426250.

[6] Dong L, Li L, Chen H, Cao Y, Lei HZAdv. Sci., 2025, 12e2403949.

[7] Olivares A O, Baker T A, Sauer R TZAnnu. Rev. Physiol., 2018, 80413.

[8] Edwards D T, LeBlanc M A, Perkins T TProc. Natl. Acad. Sci. U. S. A., 2021, 118e2015728118.

[9] Gokulu I S, Banta SZACS Synth. Biol., 2024, 132600.

[10] Wang Y Y, Ma H, Long Y T, Ying Y LZChem. Res. Chinese Universities, 2024, 40272.

[11] Ying Y L, Yang C N, Liu W, Wu X Y, Long Y TZNat. Chem., 2025, 171450.

[12] Ansah I B, Joardar S, Freedman K JZJ. Am. Chem. Soc., 2026, 14815797.

[13] Ahmed S A, Liu Y, Xiong T, Zhao Y, Xie B, Pan C, Ma W, Yu PZAnal. Chem., 2024, 968056.

[14] Brucale M, Schuler B, Samorì BZChem. Rev., 2014, 1143281.

[15] Yusko E C, Bruhn B R, Eggenberger O M, Houghtaling J, Rollings R C, Walsh N C, Nandivada S, Pindrus M, Hall A R, Sept D, Li J, Kalonia D S, Mayer MZNat. Nanotechnol., 2017, 12360.

[16] Sze J Y Y, Ivanov A P, Cass A E G, Edel J BZNat. Commun., 2017, 81552.

[17] Ma H, Wang Y, Li Y X, Xie B K, Hu Z L, Yu R J, Long Y T, Ying Y LZJ. Am. Chem. Soc., 2024, 14628014.

[18] Pan R, Wang D, Liu K, Chen H Y, Jiang DZJ. Am. Chem. Soc., 2022, 14417558.

[19] Bandara Y M N D Y, Freedman K JZJ. Am. Chem. Soc., 2024, 1463171.

[20] Wang X, Wilkinson M D, Lin X, Ren R, Willison K R, Ivanov A P, Baum J, Edel J BZChem. Sci., 2019, 11970.

[21] Fologea D, Ledden B, McNabb D S, Li JZAppl. Phys. Lett., 2007, 91053901.

[22] Guo Q, Gakhar L, Wickersham K, Francis K, Vardi-Kilshtain A, Major D T, Cheatum C M, Kohen AZBiochemistry, 2016, 552760.

[23] He L, Luo Y C, Zhang Y, Tan T F, Liu J QZChem. Res. Chinese Universities, 2025, 4195.

[24] Yoshimoto M, Yamashita T, Yamashiro TZBiotechnol. Prog., 2010, 261047.

[25] Ying Y L, Hu Z L, Zhang S, Qing Y, Fragasso A, Maglia G, Meller A, Bayley H, Dekker C, Long Y TZNat. Nanotechnol., 2022, 171136.

[26] Schmid S, Stömmer P, Dietz H, Dekker CZNat. Nanotechnol., 2021, 161244.

[27] Talaga D S, Li JZJ. Am. Chem. Soc., 2009, 1319287.

[28] Sadhu S, Soni G VZNanoscale, 2025, 1725740.

[29] Bakouei M, Abdorahimzadeh S, Taghipoor MZPhys. Chem. Chem. Phys., 2020, 2225306.

[30] Neuman K C, Nagy AZNat. Methods., 2008, 5491.

Rights & permissions

Jilin University, The Editorial Department of Chemical Research in Chinese Universities and Springer-Verlag GmbH