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Metal-controlled interdomain cooperativity in parvalbumins

Authors
Journal
Cell Calcium
0143-4160
Publisher
Elsevier
Publication Date
Volume
46
Issue
3
Identifiers
DOI: 10.1016/j.ceca.2009.07.001
Keywords
  • Thermodynamics
  • Cooperativity
  • Thermodynamic Domain
  • Structural Domain
  • Ef-Hand
  • Protein Unfolding
  • Protein Denaturation
  • Intermediate
  • Metal Binding
  • Protein Cavities
  • Protein Intrinsic Disorder
  • Hyperthermophile
  • Allergen
Disciplines
  • Biology
  • Physics

Abstract

Abstract Conformational behavior of five homologous proteins, parvalbumins (PAs) from northern pike (α and β isoforms), Baltic cod, and rat (α and β isoforms), was studied by scanning calorimetry, circular dichroism, and bis-ANS fluorescence. The mechanism of the temperature-induced denaturation of these proteins depends dramatically on both the peculiarities of their amino acid sequences and on their interaction with metal ions. For example, the pike α-PA melting can be described by two successive two-state transitions with mid-temperatures of 90 and 120 °C, suggesting the presence of two thermodynamic domains. The intermediate state populated at the end of the first transition was shown to bind Ca 2+ ions, and was characterized by the largely preserved secondary structure and increased solvent exposure of hydrophobic groups. Mg 2+- and Na +-loaded forms of pike α-PA demonstrated a single two-state transition. Therefore, the mechanism of the PA thermal denaturation is controlled by metal binding. It ranged from the absence of detectable first-order transition (apo-form of pike PA), to the two-state transition (e.g., Mg 2+- and Na +-loaded forms of pike α-PA), to the more complex mechanisms (Ca 2+-loaded PAs) involving at least one partially folded intermediate. Analysis of isolated cavities in the protein structures revealed that the interface between the CD and EF subdomains of Ca 2+-loaded pike α-PA is much more loosely packed compared with PAs manifesting single heat-sorption peak. The impairment of interactions between CD and EF subdomains may cause a loss of structural cooperativity and appearance of two separate thermodynamic domains. One more peculiar feature of pike α-PA is that depending on its interactions with metal ions, it can be an intrinsically disordered protein (apo-form), an ordered protein of mesophilic (Na +-bound state), thermophilic (Mg 2+-form), or even of the hyperthermophilic origin (Ca 2+-form).

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