Crystal versus solution structure of enzymes: NMR spectroscopy of a peptide boronic acid-serine protease complex in the crystalline state.
The effectiveness of boronic acids as inhibitors of serine proteases has been widely ascribed to the ability of the boronyl group to form a tetrahedral adduct with the active-site serine that closely mimics the putative tetrahedral intermediate or transition state formed with substrates. However, recent 15N NMR studies of alpha-lytic protease (EC 3.4.21.12) in solution have shown that some boronic acids and peptide boronic acids form adducts with the active-site histidine instead of with the serine. Such histidine-boron adducts have not thus far been reported in x-ray diffraction studies of boronic acid-serine protease complexes. Here, we report an 15N NMR study of the MeOSuc-Ala-Ala-Pro-boroPhe complex of alpha-lytic protease in the crystalline state using magic-angle spinning. Previous 15N NMR studies have shown this complex involves the formation of a histidine-boron bond in solution. The 15N NMR spectra of the crystalline complex are essentially identical to those of the complex in solution, thereby showing that the structure of this complex is the same in solution and in the crystal and that both involve formation of a histidine-boron adduct.
Department of Biochemistry, Tufts University, Boston, MA 02111.
S
S
Farr-Jones
S O
SO
Smith
C A
CA
Kettner
R G
RG
Griffin
W W
WW
Bachovchin
D000595:Amino Acid Sequence; D001897:Boronic Acids; D003460:Crystallization; D006639:Histidine; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes; D011487:Protein Conformation; D012697:Serine Endopeptidases; D012996:Solutions
D001897:Boronic Acids; D009587:Nitrogen Isotopes; D012996:Solutions; D006639:Histidine; D012697:Serine Endopeptidases; C021826:myxobacter alpha-lytic proteinase