Browsing by Author "Muddassir, Mohd"

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    Single-molecule analysis of osmolyte-mediated nanomechanical unfolding behavior of a protein domain
    (Elsevier B.V., 2023-09-16T00:00:00) Bajaj, Manish; Muddassir, Mohd; Choi, Bumjoon; Singh, Priyanka; Park, Jong Bum; Singh, Surjeet; Yadav, Manisha; Kumar, Rajesh; Eom, Kilho; Sharma, Deepak
    The small organic molecules, known as osmolytes being ubiquitously present in different cell types, affect protein folding, stability and aggregation. However, it is unknown how the osmolytes affect the nanomechanical unfolding behavior of protein domain. Here, we show the osmolyte-dependent mechanical unfolding properties of protein titin immunoglobulin-27 (I27) domain using an atomic force microscopy (AFM)-based single-molecule force spectroscopy. We found that amines and methylamines improved the mechanical stability of I27 domain, whereas polyols had no effect. Interestingly, glycine betaine (GB) or trimethylamine-N-oxide (TMAO) increased the average unfolding force of the protein domain. The kinetic parameters analyzed at single-molecule level reveal that stabilizing effect of osmolytes is due to a decrease in the unfolding rate constant of I27, which was confirmed by molecular dynamics simulations. Our study reveals different effects that diverse osmolytes have on the mechanical properties of the protein, and suggests the potential use of osmolytes in modulating the mechanical stability of proteins required for various nano-biotechnological applications. � 2023 Elsevier B.V.