Analysis of the aromatic 1H-NMR spectrum of the kringle 5 domain from human plasminogen. Evidence for a conserved kringle fold.
A kringle 5 domain fragment from human plasminogen has been investigated by 1H-NMR spectroscopy at 300 MHz and 620 MHz. The study focuses on the kringle 5 aromatic spectrum as aromatic side chains appear to mediate the binding of benzamidine. Spin-echo experiments and acid/base-titration studies in conjunction with two-dimensional double-quantum and chemical-shift-correlated spectroscopies were used to identify individual spin systems. Sequence-specific assignments of aromatic resonances are derived from direct comparison of the kringle 5 spectrum with spectra of the homologous kringle 1 and kringle 4 domains of plasminogen. As previously observed for kringles 1 and 4, the pattern we detect for Tyr9 in kringle 5 reflects a slow conformational exchange between two states in equilibrium, one in which the Tyr9 ring is freely mobile and one in which its flip dynamics are constrained. Proton Overhauser experiments in 1H2O and in 2H2O have been used to probe aromatic ring interactions and to identify residues which are part of the hydrophobic core centered at the Leu46 side chain. Overall, the data indicate a strong structural homology among the three plasminogen kringles.
Thewes
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T
Department of Chemistry, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213.
Ramesh
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V
Simplaceanu
E L
EL
Llinás
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M
D006639:Histidine; D006801:Humans; D006859:Hydrogen; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D010434:Pepsin A; D010446:Peptide Fragments / Q000302:isolation & purification; D010649:Phenylalanine; D010958:Plasminogen; D011487:Protein Conformation; D014364:Tryptophan; D014443:Tyrosine
D010446:Peptide Fragments; D014443:Tyrosine; D010649:Phenylalanine; D006639:Histidine; D006859:Hydrogen; D014364:Tryptophan; D010958:Plasminogen; D010434:Pepsin A