Subunit association and monomer structure of CINC/Gro revealed by 1H-NMR.
Rat CINC/Gro is a 72 residue chemotactic factor of neutrophils, and a member of the CXC chemokine family, that includes IL-8 and MGSA/GRO. Although the three-dimensional structure of CINC/Gro had previously been determined to be that of a dimer with 200 mM NaCl, it was shown on both ultracentrifugation analysis and 1H-NMR spectral analysis that CINC/Gro exists mainly as a monomer at a physiological concentration, similar to other proteins belonging to this family. By reducing the NaCl concentration, the equilibrium could be shifted to the monomer, making it possible to observe the monomer and dimer resonances in 1H-NMR spectra. There were no significant chemical shift changes of alpha protons in the beta sheet between the monomer and dimer, suggesting that the beta sheet structure was retained in the monomer. Instead, the chemical shift changes of a protons were significant at I18 and K21, which are located in the long loop region interacting with the alpha helix, and V59 at the beginning of the a helix, indicating structural changes in the relative positions of the alpha helix and beta sheet.
Hanzawa
H
H
Analytical and Metabolic Research Laboratories, Sankyo Co., Ltd., Shinagawa-ku, Tokyo.
Haruyama
H
H
Konishi
K
K
Watanabe
K
K
Tsurufuji
S
S
D000818:Animals; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D054360:Chemokine CXCL1; D019743:Chemokines, CXC; D002630:Chemotactic Factors / Q000737:chemistry*; D019281:Dimerization; D006133:Growth Substances / Q000737:chemistry*; D006859:Hydrogen; D006863:Hydrogen-Ion Concentration; D036341:Intercellular Signaling Peptides and Proteins; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D011485:Protein Binding; D011487:Protein Conformation; D051381:Rats; D012965:Sodium Chloride; D013696:Temperature; D014461:Ultracentrifugation / Q000379:methods
D054360:Chemokine CXCL1; D019743:Chemokines, CXC; D002630:Chemotactic Factors; C513297:Cxcl1 protein, rat; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D012965:Sodium Chloride; D006859:Hydrogen