Supramolecular Nanotube of Chaperonin GroEL: Length Control for Cellular Uptake Using Single-Ring GroEL Mutant as End-Capper

Abstract

How to modulate supramolecular protein nanotubes without sacrificing their thermodynamic stability? This challenging issue emerged with an enhanced reality since our successful development of a protein nanotube of chaperonin GroELMC as a novel ATP-responsive 1D nanocarrier because the nanotube length may potentially affect the cellular uptake efficiency. Herein, we report a molecularly engineered protein end-capper (SRMC) that firmly binds to the nanotube termini since the end-capper originates from GroEL. According to the single-ring mutation of GroEL, we obtained a single-ring version of GroEL bearing cysteine mutations (GroELCys) and modified its 14 apical cysteine residues with merocyanine (MC). Whereas SRMC self-dimerizes upon treatment with Mg(2+), we confirmed that SRMC serves as the efficient end-capper for the Mg(2+)-mediated supramolecular polymerization of GroELMC and allows for modulating the average nanotube length over a wide range from 320 to 40 nm by increasing the feed molar ratio SRMC/GroELMC up to 5.4. We also found that the nanotubes shorter than 100 nm are efficiently taken up into HEP3B cells.

Rational Design of Supramolecular Dynamic Protein Assemblies by Using a Micelle‐Assisted Activity‐Based Protein‐Labeling Technology

“…[1][2][3][4][5] Genetic engineering [6][7][8] and supramolecular chemical strategies [9][10][11][12] are two complementary technologies that are widely used for the design strategy.T he structures of protein complexes made through genetic methods have been determined to an ear-atomic resolution by using X-ray crystallography [13] and cryo-electron microscopy (cryo-EM). [1][2][3][4][5] Genetic engineering [6][7][8] and supramolecular chemical strategies [9][10][11][12] are two complementary technologies that are widely used for the design strategy.T he structures of protein complexes made through genetic methods have been determined to an ear-atomic resolution by using X-ray crystallography [13] and cryo-electron microscopy (cryo-EM).…”

Programmable Assembly of Peptide Amphiphile via Noncovalent-to-Covalent Bond Conversion

“… 1 − 6 The preparation of supramolecular polymers with monodisperse length, which could be a critical parameter for their properties, is extremely challenging due to their intrinsically dynamic nature. 7 − 9 Kinetically controlled seeded-growth and supramolecular living polymerization have been successfully used to control growth, but their implementation requires careful sample preparation and highly sophisticated molecular designs. 10 − 15 Templated growth achieved by the coassembly of supramolecular monomers with a rigid template was shown to form supramolecular nanostructures with lengths determined by the length of the template.…”

Highly ordered protein cage assemblies: A toolkit for new materials

“…The 2,700 identical major CPs (pVIII) encapsulate the genetic material, and it is capped with five copies of pIX and pVII on one apical position and five copies of pIII and pVI on the other. The proteins can display recognition peptides, which selectively bind to substrates known as phage display (Clackson, Hoogenboom, Griffiths, & Winter, 1991;Kehoe & Kay, 2005;McCafferty, Griffiths, Winter, & Chiswell, 1990;Scott & Smith, 1990;Smith & Petrenko, 1997 Sim, Miyajima, Niwa, Taguchi, and Aida (2015); Sim, Niwa, Taguchi, and Aida (2016) 1997). Additionally, the M13 phage is known to have liquid-crystalline-like behavior in highly concentrated solutions, which will be discussed in further detail in Section 3.3 (Dogic & Fraden, 2006 Stable protein 1 (SP1) is a plant-based protein commonly isolated from the aspen tree.…”