The two tryptophans of 2-microglobulin have distinct roles in function and folding and might represent two independent responses to evolutionary pressure

Sara Raimondi, Nicola Barbarini, Palma Mangione, Gennaro Esposito, Stefano Ricagno, Martino Bolognesi, Irene Zorzoli, Loredana Marchese, Cristina Soria, Riccardo Bellazzi, Maria Monti, Monica Stoppini, Mario Stefanelli, Paolo Magni, Vittorio Bellotti

    Research output: Contribution to journalArticle

    Abstract

    Background: We have recently discovered that the two tryptophans of human 2-microglobulin have distinctive roles within the structure and function of the protein. Deeply buried in the core, Trp95 is essential for folding stability, whereas Trp60, which is solvent-exposed, plays a crucial role in promoting the binding of 2-microglobulin to the heavy chain of the class I major histocompatibility complex (MHCI). We have previously shown that the thermodynamic disadvantage of having Trp60 exposed on the surface is counter-balanced by the perfect fit between it and a cavity within the MHCI heavy chain that contributes significantly to the functional stabilization of the MHCI. Therefore, based on the peculiar differences of the two tryptophans, we have analysed the evolution of 2-microglobulin with respect to these residues. Results: Having defined the 2-microglobulin protein family, we performed multiple sequence alignments and analysed the residue conservation in homologous proteins to generate a phylogenetic tree. Our results indicate that Trp60 is highly conserved, whereas some species have a Leu in position 95; the replacement of Trp95 with Leu destabilizes 2-microglobulin by 1 kcal/mol and accelerates the kinetics of unfolding. Both thermodynamic and kinetic data fit with the crystallographic structure of the Trp95Leu variant, which shows how the hydrophobic cavity of the wild-type protein is completely occupied by Trp95, but is only half filled by Leu95. Conclusions: We have established that the functional Trp60 has been present within the sequence of 2-microglobulin since the evolutionary appearance of proteins responsible for acquired immunity, whereas the structural Trp95 was selected and stabilized, most likely, for its capacity to fully occupy an internal cavity of the protein thereby creating a better stabilization of its folded state.

    Original languageEnglish (US)
    Article number159
    JournalBMC Evolutionary Biology
    Volume11
    Issue number1
    DOIs
    StatePublished - Jan 1 2011

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    tryptophan
    folding
    major histocompatibility complex
    protein
    cavity
    proteins
    thermodynamics
    stabilization
    kinetics
    sequence alignment
    immunity
    selection pressure
    replacement
    phylogenetics
    phylogeny

    ASJC Scopus subject areas

    • Ecology, Evolution, Behavior and Systematics

    Cite this

    The two tryptophans of 2-microglobulin have distinct roles in function and folding and might represent two independent responses to evolutionary pressure. / Raimondi, Sara; Barbarini, Nicola; Mangione, Palma; Esposito, Gennaro; Ricagno, Stefano; Bolognesi, Martino; Zorzoli, Irene; Marchese, Loredana; Soria, Cristina; Bellazzi, Riccardo; Monti, Maria; Stoppini, Monica; Stefanelli, Mario; Magni, Paolo; Bellotti, Vittorio.

    In: BMC Evolutionary Biology, Vol. 11, No. 1, 159, 01.01.2011.

    Research output: Contribution to journalArticle

    Raimondi, S, Barbarini, N, Mangione, P, Esposito, G, Ricagno, S, Bolognesi, M, Zorzoli, I, Marchese, L, Soria, C, Bellazzi, R, Monti, M, Stoppini, M, Stefanelli, M, Magni, P & Bellotti, V 2011, 'The two tryptophans of 2-microglobulin have distinct roles in function and folding and might represent two independent responses to evolutionary pressure', BMC Evolutionary Biology, vol. 11, no. 1, 159. https://doi.org/10.1186/1471-2148-11-159
    Raimondi, Sara ; Barbarini, Nicola ; Mangione, Palma ; Esposito, Gennaro ; Ricagno, Stefano ; Bolognesi, Martino ; Zorzoli, Irene ; Marchese, Loredana ; Soria, Cristina ; Bellazzi, Riccardo ; Monti, Maria ; Stoppini, Monica ; Stefanelli, Mario ; Magni, Paolo ; Bellotti, Vittorio. / The two tryptophans of 2-microglobulin have distinct roles in function and folding and might represent two independent responses to evolutionary pressure. In: BMC Evolutionary Biology. 2011 ; Vol. 11, No. 1.
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    abstract = "Background: We have recently discovered that the two tryptophans of human 2-microglobulin have distinctive roles within the structure and function of the protein. Deeply buried in the core, Trp95 is essential for folding stability, whereas Trp60, which is solvent-exposed, plays a crucial role in promoting the binding of 2-microglobulin to the heavy chain of the class I major histocompatibility complex (MHCI). We have previously shown that the thermodynamic disadvantage of having Trp60 exposed on the surface is counter-balanced by the perfect fit between it and a cavity within the MHCI heavy chain that contributes significantly to the functional stabilization of the MHCI. Therefore, based on the peculiar differences of the two tryptophans, we have analysed the evolution of 2-microglobulin with respect to these residues. Results: Having defined the 2-microglobulin protein family, we performed multiple sequence alignments and analysed the residue conservation in homologous proteins to generate a phylogenetic tree. Our results indicate that Trp60 is highly conserved, whereas some species have a Leu in position 95; the replacement of Trp95 with Leu destabilizes 2-microglobulin by 1 kcal/mol and accelerates the kinetics of unfolding. Both thermodynamic and kinetic data fit with the crystallographic structure of the Trp95Leu variant, which shows how the hydrophobic cavity of the wild-type protein is completely occupied by Trp95, but is only half filled by Leu95. Conclusions: We have established that the functional Trp60 has been present within the sequence of 2-microglobulin since the evolutionary appearance of proteins responsible for acquired immunity, whereas the structural Trp95 was selected and stabilized, most likely, for its capacity to fully occupy an internal cavity of the protein thereby creating a better stabilization of its folded state.",
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    AU - Raimondi, Sara

    AU - Barbarini, Nicola

    AU - Mangione, Palma

    AU - Esposito, Gennaro

    AU - Ricagno, Stefano

    AU - Bolognesi, Martino

    AU - Zorzoli, Irene

    AU - Marchese, Loredana

    AU - Soria, Cristina

    AU - Bellazzi, Riccardo

    AU - Monti, Maria

    AU - Stoppini, Monica

    AU - Stefanelli, Mario

    AU - Magni, Paolo

    AU - Bellotti, Vittorio

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    N2 - Background: We have recently discovered that the two tryptophans of human 2-microglobulin have distinctive roles within the structure and function of the protein. Deeply buried in the core, Trp95 is essential for folding stability, whereas Trp60, which is solvent-exposed, plays a crucial role in promoting the binding of 2-microglobulin to the heavy chain of the class I major histocompatibility complex (MHCI). We have previously shown that the thermodynamic disadvantage of having Trp60 exposed on the surface is counter-balanced by the perfect fit between it and a cavity within the MHCI heavy chain that contributes significantly to the functional stabilization of the MHCI. Therefore, based on the peculiar differences of the two tryptophans, we have analysed the evolution of 2-microglobulin with respect to these residues. Results: Having defined the 2-microglobulin protein family, we performed multiple sequence alignments and analysed the residue conservation in homologous proteins to generate a phylogenetic tree. Our results indicate that Trp60 is highly conserved, whereas some species have a Leu in position 95; the replacement of Trp95 with Leu destabilizes 2-microglobulin by 1 kcal/mol and accelerates the kinetics of unfolding. Both thermodynamic and kinetic data fit with the crystallographic structure of the Trp95Leu variant, which shows how the hydrophobic cavity of the wild-type protein is completely occupied by Trp95, but is only half filled by Leu95. Conclusions: We have established that the functional Trp60 has been present within the sequence of 2-microglobulin since the evolutionary appearance of proteins responsible for acquired immunity, whereas the structural Trp95 was selected and stabilized, most likely, for its capacity to fully occupy an internal cavity of the protein thereby creating a better stabilization of its folded state.

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