Laboratory experiment demonstrates reversible splitting of tissue thromboplastin into inactive parts, indicating that protein and phospholipid fractions are both required for clotting.
Key Points
To determine whether tissue thromboplastin can be dissociated into separable constituent parts and subsequently reassembled to regain its biological activity.
Treated tissue thromboplastin with the detergent sodium deoxycholate (DOC) to dissociate the complex.
Separated the dissociated products into fraction A and fraction B using gel filtration chromatography in the presence of DOC.
Recombined the separated fractions and dialyzed or processed them to remove DOC, assessing for structural reaggregation and clotting activity.
Sodium deoxycholate treatment split tissue thromboplastin into two inactive components: fraction A, consisting predominantly of protein, and fraction B, consisting predominantly of phospholipid.
Recombining fractions A and B followed by removal of sodium deoxycholate induced reaggregation and fully restored thromboplastic activity.