High-resolution NMR studies of fibrinogen-like peptides in solution: Structure of a thrombin-bound peptide corresponding to residues 7-16 of the A. alpha. chain of human fibrinogen
- Cornell Univ., Ithaca, NY (USA)
The interaction of the following human fibrinogen-like peptides with bovine thrombin was studied by one- and two-dimensional NMR techniques in aqueous solution: acetyl-Phe(8)-Leu(9)-Ala(10)-Glu(11)-Gly(12)-Gly(13)-Gly(14)-Val(15)-Arg(16)-Gly(17)-Pro(18)-NHMe (F6), acetyl-Phe-Leu-Ala-Glu-Gly-Gly-Gly-Val-Arg(16) (tF6), acetyl-Asp(7)-Phe-Leu-Ala-Glu-Gly-Gly-Gly-Val-Arg(16)-Gly(17)-Pro-Arg(19)-Val(20)-NHMe (F8), and acetyl-Asp-Phe-Leu-Ala-Glu-Gly-Gly-Gly-Val-Arg(16) (tF8). At pH 5.3 and 25{degree}C, the Arg(16)-Gly(17) peptide bonds in both F6 and F8 were cleaved instantaneously in the presence of 0.5 mM thrombin, producing truncated peptides tF6 and tF8 and other peptide fragments. On the basis of observations of line broadening, thrombin was found to bind to the cleavage products, tF6 and tF8, of peptides F6 and F8. Transferred NOE (TRNOE) measurements were made of the complexes between thrombin and peptides tF6 and tF8. Medium- and long-range NOE interactions were found between the NH proton of Asp(7) and the C{sup {beta}}H protons of Ala(10), between the C{sup {alpha}}H proton of Glu(11) and the NH proton of Gly(13), and between the ring protons of Phe(8) and the C{sup {alpha}}H protons of Gly(14) and the C{sup {gamma}}H protons of Val(15). Sets of structures of the decapeptide tF8 were deduced by use of distance geometry calculations based on sequential and medium- and long-range TRNOEs from the thrombin-bound peptide. These results provide an explanation for the observations that Asp(7), Phe(8), and Gly(12) are strongly conserved in mammalian fibrinogens and that the mutations of Asp(7) to Asn(7) and of Gly(12) to Val(12) result in delayed release of fibrinopeptide A, producing human bleeding disorders.
- OSTI ID:
- 5605574
- Journal Information:
- Biochemistry; (USA), Journal Name: Biochemistry; (USA) Vol. 28:7; ISSN 0006-2960; ISSN BICHA
- Country of Publication:
- United States
- Language:
- English
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High-resolution NMR studies of fibrinogen-like peptides in solution: resonance assignments and conformational analysis of residues 1-23 of the A. cap alpha. chain of human fibrinogen
High-resolution NMR studies of fibrinogen-like peptides in solution: Interaction of thrombin with residues 1-23 of the A. alpha. chain of human fibrinogen
Related Subjects
62 RADIOLOGY AND NUCLEAR MEDICINE
AMIDES
AMINO ACIDS
ANIMALS
ASPARAGINE
ASPARTIC ACID
BARYONS
BLOOD COAGULATION FACTORS
CARBOXYLIC ACIDS
CATTLE
CHROMATOGRAPHY
COAGULANTS
DOMESTIC ANIMALS
DRUGS
ELEMENTARY PARTICLES
FERMIONS
FIBRINOGEN
GLOBULINS
GLYCINE
HADRONS
HEMATOLOGIC AGENTS
HEMOSTATICS
LEUCINE
LIQUID COLUMN CHROMATOGRAPHY
MAGNETIC RESONANCE
MAMMALS
MAN
MOLECULAR STRUCTURE
NUCLEAR MAGNETIC RESONANCE
NUCLEONS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PEPTIDES
PRIMATES
PROTEINS
PROTONS
RESONANCE
RUMINANTS
SEPARATION PROCESSES
VALINE
VERTEBRATES