title: Molecular interaction studies of hemostasis: fibrinogen ligand-human platelet receptor interactions.
abstract: The interactions between fibrinogen ligands and platelet receptor alpha(IIb)beta(3) were studied under physiological conditions by atomic force microscopy (AFM). Two linear peptide sequences in fibrinogen, RGD and HHLGGAKQAGDV, play central roles in the regulation of hemostasis and thrombosis by facilitating adhesion and aggregation of platelets. In order to measure the interactions (i.e., debonding force), oligopeptides, GSSSGaaa, where aaa is -RGDSPA or -HHLGGAKQAGDV, were synthesized and grafted on to the surface of AFM probe tips. The interaction forces between a peptide-modified AFM probe tip and platelet surface were determined from pN to nN levels using AFM force measurements. Our results show that the zero kinetic off-rate, K(off)(0), for RGDSPA is significantly smaller than that for HHLGGAKQAGDV, under the consideration of flexible receptor surfaces. From our analysis, the K(off)(0), the single molecular binding energy E(b), and the transition state x(b), were extracted from the data, and estimated to be 1.53s(-1), -2.64x10(-20)J and 1.03A for the RGD-alpha(IIb)beta(3) system, and 47.58s(-1), 2.67x10(-20), 1.09A for the HHLGGAKQAGDV-alpha(IIb)beta(3) system, respectively.
authors.affiliation: Department of Biomedical Engineering, Case Western Reserve University, 10900 Eculid Avenue, Wickenden Building, Cleveland, OH 44106-7207, USA.
authors.forename: Imshik
authors.initials: I
authors.lastname: Lee
authors.forename: Roger E
authors.initials: RE
authors.lastname: Marchant
mesh_terms: D000595:Amino Acid Sequence; D001792:Blood Platelets / Q000378:metabolism*; D005340:Fibrinogen / Q000737:chemistry* / Q000378:metabolism*; D006487:Hemostasis; D006801:Humans; D008024:Ligands; D018625:Microscopy, Atomic Force; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000378:metabolism*
chemical_list: D008024:Ligands; D010455:Peptides; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D005340:Fibrinogen