title: Multidomain structure of a recombinant streptokinase. A differential scanning calorimetry study.
abstract: The temperature dependence of the heat capacity function of a recombinant streptokinase (rSK) has been studied by high-sensitivity differential scanning microcalorimetry and circular dichroism as a function of pH in low- and high-ionic strength buffers. At low ionic strength it is found that this protein, between pH 7 and 10, undergoes four reversible and independent two-state transitions during its unfolding, suggesting the existence of four domains in the native structure of the protein. This result reconciles previous conflicting reports about the number of domains of this protein obtained by differential scanning calorimetry and small-angle X-ray scattering. The number of two-state transitions decreases when the pH of the medium is decreased, without noticeable changes in its circular dichroism spectrum. A plausible localization of the four domains in the streptokinase sequences is proposed and their thermodynamic parameters are given. Increase of ionic strength to 200 mM NaCl affects positively the protein stability and confirms the existence of four reversible two-state transitions. Above 200 mM NaCl the protein stability decreases, resulting in low percentage of reversibility, and even irreversible transitions.
authors.affiliation: Centro de Ingeniería Genética y Biotecnología, Havana, Cuba. evalproc@ta.ciqb.edu.cu
authors.forename: A
authors.initials: A
authors.lastname: Beldarraín
authors.forename: J L
authors.initials: JL
authors.lastname: López-Lacomba
authors.forename: V P
authors.initials: VP
authors.lastname: Kutyshenko
authors.forename: R
authors.initials: R
authors.lastname: Serrano
authors.forename: M
authors.initials: M
authors.lastname: Cortijo
mesh_terms: D002152:Calorimetry, Differential Scanning; D002942:Circular Dichroism; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D009154:Mutation; D011487:Protein Conformation; D011489:Protein Denaturation; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D012965:Sodium Chloride / Q000737:chemistry; D013300:Streptokinase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D012965:Sodium Chloride; D013300:Streptokinase