Relative stability of human centrins and its relationship to calcium binding.
Centrins are calcium binding proteins that belong to the EF-hand superfamily with diverse biological functions. Herein we present the first systematic study that establishes the relative stability of related centrins via complementary biophysical techniques. Our results define the stepwise molecular behavior of human centrins by two-dimensional infrared (2D IR) correlation spectroscopy, the change in heat capacity and enthalpy of denaturation by differential scanning calorimetry, and the relative stability of the helical regions of centrins by circular dichroism. More importantly, 2D IR correlation spectroscopy provides unique information about the similarities and differences in dynamics between these related proteins. The thermally induced molecular behavior of human centrins can be used to predict biological target interactions that have a relative dependence on calcium affinity. This information is essential for understanding why certain isoforms may be used to rescue a phenotype and therefore also for explaining the different functions these proteins may have in vivo. Furthermore, this comparative approach can be applied to the study of recombinant therapeutic protein candidates for the treatment of disease states.
Protein Research Center, University of Puerto Rico, Mayagüez Campus, Mayagüez, Puerto Rico 00681-9019, USA. belinda@hpcf.upr.edu
Belinda
B
Pastrana-Ríos
Myrna
M
Reyes
Jessica
J
De Orbeta
Verónica
V
Meza
Daniel
D
Narváez
Ana María
AM
Gómez
Aslin
A
Rodríguez Nassif
Ruth
R
Almodovar
Adalberto
A
Díaz Casas
José
J
Robles
Ana María
AM
Ortiz
Lizbeth
L
Irizarry
Melissa
M
Campbell
Mara
M
Colón
D000595:Amino Acid Sequence; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D002152:Calorimetry, Differential Scanning; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000378:metabolism; D002942:Circular Dichroism; D006801:Humans; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011489:Protein Denaturation; D017550:Spectroscopy, Fourier Transform Infrared; D013057:Spectrum Analysis / Q000379:methods
C580665:CETN1 protein, human; C472752:CETN2 protein, human; C107745:CETN3 protein, human; D002135:Calcium-Binding Proteins; D018797:Cell Cycle Proteins; D002118:Calcium