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

Journal Article · Tue Apr 04 00:00:00 EDT 1989 · Biochemistry; (USA)

DOI:https://doi.org/10.1021/bi00433a053· OSTI ID:5605574

Ni, F; Meinwald, Y C; Vasquez, M; Scheraga, H A ^[1]

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1. 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), Vol. 28:7; ISSN 0006-2960
• Country of Publication: United States
• Language: English

Related Subjects

62 RADIOLOGY AND NUCLEAR MEDICINE
FIBRINOGEN
MOLECULAR STRUCTURE
PEPTIDES
NUCLEAR MAGNETIC RESONANCE
ASPARAGINE
ASPARTIC ACID
CATTLE
GLYCINE
LEUCINE
LIQUID COLUMN CHROMATOGRAPHY
MAN
PROTONS
VALINE
AMIDES
AMINO ACIDS
ANIMALS
BARYONS
BLOOD COAGULATION FACTORS
CARBOXYLIC ACIDS
CHROMATOGRAPHY
COAGULANTS
DOMESTIC ANIMALS
DRUGS
ELEMENTARY PARTICLES
FERMIONS
GLOBULINS
HADRONS
HEMATOLOGIC AGENTS
HEMOSTATICS
MAGNETIC RESONANCE
MAMMALS
NUCLEONS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PRIMATES
PROTEINS
RESONANCE
RUMINANTS
SEPARATION PROCESSES
VERTEBRATES
550601* - Medicine- Unsealed Radionuclides in Diagnostics