Purification of salmon thrombin and its potential as an alternative to mammalian thrombins in fibrin sealants.
A method to produce highly purified thrombin from salmon blood is described, and a series of biochemical, cell biologic, and biophysical assays demonstrate the functional similarities and some differences between salmon and human thrombins. Salmon thrombin with specific activity greater than 1000 units/mg total protein can be prepared by modifications of the methods used for purification of human thrombin. Using a synthetic substrate based on the human fibrinogen A-alpha polypeptide sequence as an indicator of enzymatic activity, salmon and human thrombin preparations contain similar specific activities per mass of purified protein. Salmon thrombin activates human fibrinogen and initiates the formation of fibrin clots whose structure and rheologic properties are indistinguishable from those of human fibrin clotted by human thrombin. Salmon thrombin also activates human platelets. Approximately 10 times higher activities are needed for the same rate of platelet aggregation compared to human thrombin, and some aspects of platelet activation, most notably phosphatidylserine exposure, are diminished relative to the effects of human thrombin. This latter finding suggests that salmon thrombin may not activate all of the receptors that are targets of human thrombin, although it does appear to activate signals that are sufficient to produce normal rates of activation and aggregation as measured by conventional aggregometry. Together with the recent purification of salmon fibrinogen and its application in mammalian wound healing, the availability of salmon thrombin allows the formulation of biological sealants devoid of any exogenous mammalian proteins and so may aid the design of materials with increased safety from infectious disease transmission.
Michaud
Sarah E
SE
Hematology Division, Brigham and Women's Hospital, LMRC #302, 221 Longwood Ave, Boston, MA 02115, USA.
Wang
Louise Z
LZ
Korde
Neha
N
Bucki
Robert
R
Randhawa
Paramjeet K
PK
Pastore
Jennifer J
JJ
Falet
Hervé
H
Hoffmeister
Karin
K
Kuuse
Reet
R
Uibo
Raivo
R
Herod
Julia
J
Sawyer
Evelyn
E
Janmey
Paul A
PA
D000818:Animals; D000910:Antibodies, Heterophile / Q000276:immunology; D003429:Cross Reactions; D004341:Drug Evaluation; D015718:Fibrin Tissue Adhesive / Q000737:chemistry; D005340:Fibrinogen / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D015539:Platelet Activation / Q000187:drug effects; D012474:Salmon; D013917:Thrombin / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology*
D000910:Antibodies, Heterophile; D015718:Fibrin Tissue Adhesive; D005340:Fibrinogen; D013917:Thrombin