Multiplexed silicon photonic sensor arrays enable facile characterization of coagulation protein binding to nanodiscs with variable lipid content

J Biol Chem. 2017 Sep 29;292(39):16249-16256. doi: 10.1074/jbc.M117.800938. Epub 2017 Aug 11.


Interactions of soluble proteins with the cell membrane are critical within the blood coagulation cascade. Of particular interest are the interactions of γ-carboxyglutamic acid-rich domain-containing clotting proteins with lipids. Variability among conventional analytical methods presents challenges for comparing clotting protein-lipid interactions. Most previous studies have investigated only a single clotting protein and lipid composition and have yielded widely different binding constants. Herein, we demonstrate that a combination of lipid bilayer nanodiscs and a multiplexed silicon photonic analysis technology enables high-throughput probing of many protein-lipid interactions among blood-clotting proteins. This approach allowed direct comparison of the binding constants of prothrombin, factor X, activated factor VII, and activated protein C to seven different binary lipid compositions. In a single experiment, the binding constants of one protein interacting with all lipid compositions were simultaneously determined. A simple surface regeneration then facilitated similar binding measurements for three other coagulation proteins. As expected, our results indicated that all proteins exhibit tighter binding (lower Kd ) as the proportion of anionic lipid increases. Interestingly, at high proportions of phosphatidylserine, the Kd values of all four proteins began to converge. We also found that although koff values for all four proteins followed trends similar to those observed for the Kd values, the variation among the proteins was much lower, indicating that much of the variation came from the kinetic binding (kon) of the proteins. These findings indicate that the combination of silicon photonic microring resonator arrays and nanodiscs enables rapid interrogation of biomolecular binding interactions at model cell membrane interfaces.

Keywords: coagulation factors; factor VIII (FVIII); lipid; membrane bilayer; nanodisc; phosphatidic acid; phosphatidylserine; protein–lipid interaction; prothrombin; silicon photonic microring resonators.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Factor VIIa / chemistry
  • Factor VIIa / genetics
  • Factor VIIa / metabolism*
  • Factor X / chemistry
  • Factor X / metabolism*
  • High-Throughput Screening Assays
  • Humans
  • Kinetics
  • Lipid Bilayers / chemistry
  • Lipid Bilayers / metabolism
  • Nanostructures / chemistry
  • Optical Phenomena
  • Phosphatidic Acids / chemistry
  • Phosphatidic Acids / metabolism*
  • Phosphatidylcholines / chemistry
  • Phosphatidylcholines / metabolism*
  • Phosphatidylserines / chemistry
  • Phosphatidylserines / metabolism*
  • Protein Array Analysis
  • Protein C / chemistry
  • Protein C / metabolism*
  • Prothrombin / chemistry
  • Prothrombin / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Silicon / chemistry


  • 1-palmitoyl-2-oleoyl-glycero-3-phosphatidic acid
  • Lipid Bilayers
  • Phosphatidic Acids
  • Phosphatidylcholines
  • Phosphatidylserines
  • Protein C
  • Recombinant Proteins
  • 1-palmitoyl-2-oleoylglycero-3-phosphoserine
  • Prothrombin
  • Factor X
  • Factor VIIa
  • 1-palmitoyl-2-oleoylphosphatidylcholine
  • Silicon