Dynamics of hydrogen-deuterium exchange in Chlamydomonas centrin.
Chlamydomonas reinhardtii centrin is a 169-amino acid residue calcium binding protein belonging to the EF-hand protein superfamily. Centrin is associated with the microtubule organizing center (MTOC) in all eukaryotes, and in Chlamydomonas, centrin is a component of the flagellar basal body apparatus. Recombinant full-length centrin, calmodulin, and terminal domain fragments [Ccen-N (residues 1-94) and Ccen-C (residues 99-169)] were used to examine hydrogen-deuterium (H --> D) exchange dynamics using combined attenuated total reflectance (ATR) Fourier transform-infrared (FT-IR) spectroscopy, curve fit, and two-dimensional correlation analysis. Analysis of the Ccen-N and Ccen-C fragments allowed separation of domain specific solvent exchange events and together with analysis of the full-length proteins provides novel insight into domain accessibility to the aqueous environment and the internal dynamics of the protein.
Ortiz
Mildred
M
Department of Chemistry and Center for Protein Structure Function and Dynamics, University of Puerto Rico, Mayagüez Campus, P.O. Box 9019, Mayagüez, Puerto Rico 00681-9019.
Sanoguet
Zuleika
Z
Hu
Haitao
H
Chazin
Walter J
WJ
McMurray
Cynthia T
CT
McMurray
Cynthia
C
Salisbury
Jeffrey L
JL
Pastrana-Rios
Belinda
B
D000595:Amino Acid Sequence; D000818:Animals; D002135:Calcium-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D002147:Calmodulin / Q000737:chemistry / Q000378:metabolism; D016825:Chlamydomonas reinhardtii / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry* / Q000378:metabolism*; D003285:Contractile Proteins / Q000737:chemistry / Q000378:metabolism; D041961:Deuterium Exchange Measurement / Q000379:methods*; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D012997:Solvents; D017550:Spectroscopy, Fourier Transform Infrared / Q000706:statistics & numerical data; D018709:Statistics, Nonparametric; D013816:Thermodynamics
D002135:Calcium-Binding Proteins; D002147:Calmodulin; D002868:Chromosomal Proteins, Non-Histone; D003285:Contractile Proteins; D010446:Peptide Fragments; D012997:Solvents; C056608:caltractin