title: Influence of chemical modification of tryptophan residues on the properties of human antithrombin III.
abstract: According to the reaction conditions selected, chemical modification of tryptophan residues in antithrombin III by dimethyl (2-hydroxy-5 nitrobenzyl) sulfonium bromide (HNBSB) generated products with similar levels of modification (equivalent to 0.9 mole 2-hydroxy-5-nitrobenzyl (HNB) incorporated/mole of antithrombin III) but with high or low affinity for heparin. These products were subjected to digestion by cyanogen bromide and shown to be modified equivalently in fragment II containing Trp 189 and Trp 225 and fragment III containing Trp 49. The molar level of incorporation of HNB into these fragments was similar in the high and low affinity forms. Both high and low affinity forms showed loss of heparin cofactor activity. A recovery of heparin cofactor activity towards coagulation factor Xa was observed upon prolonged storage of low affinity forms at -70 degrees C. It is considered that the loss of high affinity for heparin upon modification of antithrombin III arises from change or stabilization of conformation associated with tryptophan modification and is not a singular property of modification of Trp 49.
authors.affiliation: Thrombosis Research Unit, King's College School of Medicine & Dentistry, London, United Kingdom.
authors.forename: N
authors.initials: N
authors.lastname: Shah
authors.forename: M F
authors.initials: MF
authors.lastname: Scully
authors.forename: V
authors.initials: V
authors.lastname: Ellis
authors.forename: V V
authors.initials: VV
authors.lastname: Kakkar
mesh_terms: D000990:Antithrombin III / Q000378:metabolism*; D004355:Drug Stability; D004356:Drug Storage; D015951:Factor Xa / Q000378:metabolism; D006493:Heparin / Q000378:metabolism; D006801:Humans; D010446:Peptide Fragments / Q000032:analysis; D011485:Protein Binding; D011487:Protein Conformation; D014364:Tryptophan / Q000031:analogs & derivatives / Q000502:physiology*
chemical_list: D010446:Peptide Fragments; D014364:Tryptophan; D000990:Antithrombin III; D006493:Heparin; D015951:Factor Xa