Antithrombin III Basel. Identification of a Pro-Leu substitution in a hereditary abnormal antithrombin with impaired heparin cofactor activity.
Antithrombin III Basel is a hereditary abnormal antithrombin with normal progressive inhibition activity (normal reactive site) and reduced heparin cofactor activity (impaired heparin binding site). Structures of antithrombin III Basel and normal antithrombin III isolated from the same patient were compared by peptide mapping using the dimethylaminoazobenzene isothiocyanate precolumn derivatization technique. Of the approximately 50 tryptic peptides of normal and abnormal antithrombin III, one peptide comprising residues 40-46 had a different retention time in reversed-phase high performance liquid chromatography. The amino acid sequence of the peptide from antithrombin III Basel had a single substitution of Pro (normal) by Leu (abnormal) at position 41. This substitution is close to an Arg (residue 47) and a Trp (residue 49) which have previously been shown to be critical for heparin binding by antithrombin III. Although additional amino acid substitutions in antithrombin III Basel cannot be ruled out, this Pro-Leu replacement could cause a conformational change by increasing both the helical structure and the hydrophobicity around residue 41. These data suggest that: (i) the heparin binding site of antithrombin III encompasses the region containing residues 41, 47, and 49; and (ii) the impaired heparin cofactor activity of antithrombin III Basel is likely due to a conformational change of the heparin binding site induced by the Pro-Leu substitution at position 41.
Chang
J Y
JY
Tran
T H
TH
D000595:Amino Acid Sequence; D000990:Antithrombin III / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D017879:Isothiocyanates; D007930:Leucine / Q000032:analysis*; D010446:Peptide Fragments / Q000032:analysis; D011392:Proline / Q000032:analysis*; D013329:Structure-Activity Relationship; D013861:Thiocyanates; D013867:Thiohydantoins; D014357:Trypsin / Q000378:metabolism; D004124:p-Dimethylaminoazobenzene / Q000031:analogs & derivatives
D017879:Isothiocyanates; D010446:Peptide Fragments; D013861:Thiocyanates; D013867:Thiohydantoins; C047699:antithrombin III Basel; C022927:DABTH; C012926:4-(N,N-dimethylaminoazobenzene)-4'-isothiocyanate; D000990:Antithrombin III; D011392:Proline; D004124:p-Dimethylaminoazobenzene; D014357:Trypsin; D007930:Leucine