Interaction of heparin with annexin V.
The energetics and kinetics of the interaction of heparin with the Ca2+ and phospholipid binding protein annexin V, was examined and the minimum oligosaccharide sequence within heparin that binds annexin V was identified. Affinity chromatography studies confirmed the Ca2+ dependence of this binding interaction. Analysis of the data obtained from surface plasmon resonance afforded a Kd of approximately 21 nM for the interaction of annexin V with end-chain immobilized heparin and a Kd of approximately 49 nM for the interaction with end-chain immobilized heparan sulfate. Isothermal titration calorimetry showed the minimum annexin V binding oligosaccharide sequence within heparin corresponds to an octasaccharide sequence. The Kd of a heparin octasaccharide binding to annexin V was approximately 1 microM with a binding stoichiometry of 1:1.
Capila
I
I
Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City 52242, USA.
VanderNoot
V A
VA
Mealy
T R
TR
Seaton
B A
BA
Linhardt
R J
RJ
D000818:Animals; D017304:Annexin A5 / Q000378:metabolism*; D002151:Calorimetry; D002240:Carbohydrate Sequence; D002846:Chromatography, Affinity; D006025:Glycosaminoglycans / Q000378:metabolism; D006493:Heparin / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D017186:Titrimetry
D017304:Annexin A5; D006025:Glycosaminoglycans; D011994:Recombinant Proteins; D006493:Heparin