• Login
    View Item 
    •   DSpace@RPI Home
    • Rensselaer Libraries
    • RPI Theses Online (Complete)
    • View Item
    •   DSpace@RPI Home
    • Rensselaer Libraries
    • RPI Theses Online (Complete)
    • View Item
    JavaScript is disabled for your browser. Some features of this site may not work without it.

    Molecular determinants of the thermal expansivity of proteins and volumetric changes upon protein denaturation

    Author
    Pandharipande, Pranav
    View/Open
    177378_Pandharipande_rpi_0185E_10866.pdf (2.225Mb)
    Other Contributors
    Makhatadze, George I.; Cramer, Steven M.; Karande, Pankaj; Plawsky, Joel L., 1957-;
    Date Issued
    2016-05
    Subject
    Chemical and biological engineering
    Degree
    PhD;
    Terms of Use
    This electronic version is a licensed copy owned by Rensselaer Polytechnic Institute, Troy, NY. Copyright of original work retained by author.;
    Metadata
    Show full item record
    URI
    https://hdl.handle.net/20.500.13015/1716
    Abstract
    In this thesis document, I have extensively discussed a variety of molecular determinants of native state expansivity, established how the thermal expansivity obtained using PPC can be used as a probe to study protein compaction, characterized the volumetric properties of the secondary structural elements such as α-helix and β- sheet using PPC, probed the molecular determinants of the change in volume upon protein unfolding (ΔVvoid and ΔVhydration) and finally, shed light on the protein volume paradox resolution and its biological implications for life at extreme environmental conditions.; Engineering proteins with high stability (ΔG), improved efficiency (kcat/Km), and better specificity (Kd) requires understanding of their thermodynamics of formation, stabilization, and function. Since thermodynamics of proteins has been conventionally studied in terms of response to thermal or chemical denaturants, there is increasing interest in understanding the effect of relatively less explored hydrostatic pressure (P). The pressure dependence of stability (ΔG) is defined by the change in volume upon denaturation, ΔV = VU –VN = (∂ΔG/∂P)T. Understanding the individual factors in total change in volume upon protein unfolding poses a fundamental problem, is highly biologically relevant in terms of life in the deep ocean as well as will allow us to develop rational approaches to engineer proteins with high pressure tolerance.; Temperature dependence of change in volume upon denaturation is defined by the changes in thermal expansivity (ΔE), ΔE = (∂ΔV/∂T) P = EU –EN. The knowledge of EN and EU, is as much important as knowledge VU and VN, as they provide insights related to hydration. PPC allows experimental measurement of the thermal expansion coefficient, α = E/V, of a protein in the native, αN (T), and unfolded, αU (T), states as a function of temperature. Analysis of the thermal expansion coefficient profile, obtained over a broad temperature range, provides us the change in volume upon protein unfolding (ΔV). Thus, not only can PPC thermally denature the protein but it also can obtain thermal expansion coefficients while doing so, two state analysis of which provides volumetric changes upon protein denaturation. Hence, PPC serves a dual purpose of measuring thermostability as well as pressure stability.;
    Description
    May 2016; School of Engineering
    Department
    Dept. of Chemical and Biological Engineering;
    Publisher
    Rensselaer Polytechnic Institute, Troy, NY
    Relationships
    Rensselaer Theses and Dissertations Online Collection;
    Access
    Restricted to current Rensselaer faculty, staff and students. Access inquiries may be directed to the Rensselaer Libraries.;
    Collections
    • RPI Theses Online (Complete)

    Browse

    All of DSpace@RPICommunities & CollectionsBy Issue DateAuthorsTitlesSubjectsThis CollectionBy Issue DateAuthorsTitlesSubjects

    My Account

    Login

    DSpace software copyright © 2002-2022  DuraSpace
    Contact Us | Send Feedback
    DSpace Express is a service operated by 
    Atmire NV