The impact of folding modes and deuteration on the atomic resolution structure of hen egg-white lysozyme

Abstract

The biological function of a protein is intimately related to its structure and dynamics, which in turn are determined by the way in which it has been folded. In vitro refolding is commonly used for the recovery of recombinant proteins that are expressed in the form of inclusion bodies and is of central interest in terms of the folding pathways that occur in vivo. Here, biophysical data are reported for in vitro-refolded hydrogenated hen egg-white lysozyme, in combination with atomic resolution X-ray diffraction analyses, which allowed detailed comparisons with native hydrogenated and refolded perdeuterated lysozyme. Distinct folding modes are observed for the hydrogenated and perdeuterated refolded variants, which are determined by conformational changes to the backbone structure of the Lys97–Gly104 flexible loop. Surprisingly, the structure of the refolded perdeuterated protein is closer to that of native lysozyme than that of the refolded hydrogenated protein. These structural differences suggest that the observed decreases in thermal stability and enzymatic activity in the refolded perdeuterated and hydrogenated proteins are consequences of the macromolecular deuteration effect and of distinct folding dynamics, respectively. These results are discussed in the context of both in vitro and in vivo folding, as well as of lysozyme amyloidogenesis.

The In Vitro Antimicrobial and Antibiofilm Activities of Lysozyme against Gram-Positive Bacteria

“…It catalyzes the breaking of β -1,4-glycosidic bonds of peptidoglycan in bacterial cell walls [ 1 , 2 ] and exerts broad-spectrum antibacterial activities against Gram-negative bacteria (GNB) and Gram-positive bacteria (GPB) in vitro [ 2 ]. LYS is present not only in human [ 3 ] but also in the egg whites of most avians [ 4 ]. Additionally, LYS also has many other biological properties, such as antimicrobial [ 2 ], anti-inflammatory [ 5 ], antitumor [ 6 , 7 ], and antiviral [ 1 ] activities.…”

The effect of novel antihypertensive drug valsartan on lysozyme aggregation: A combined in situ and in silico study

“…10 D, VAL binds directly to ALA 11 in BS-2 via hydrogen bonds in a 1:1 M ratio to HEWL. Thus, the C-helix becomes less packed and less stable, leading to unfolding [ 66 ]. Additionally, interactions between ALA 11 and VAL may indirectly affect HIS 15 , THR 89 , and VAL 92 , altering the folding structure of HEWL.…”

Investigation of the structure of protein–polymer conjugates in solution reveals the impact of protein deuteration and the size of the polymer on its thermal stability

“…It can impact the protein's structural features, including hydrogen bonds and interactions with hydrophobic regions. (Nichols et al, 2020;Ramos et al, 2021;Tempra et al, 2023). This substitution has potential benefits, notably in improving the pharmacokinetics and reducing the toxicity of drugs, thereby minimizing adverse effects (Belete, 2022;Maria et al, 2023).…”