Characterization and purification of tissue plasminogen activator and its binding to the surface of cerebellar neutrons
Early studies directed at understanding the possible role of the fibrinolytic system in the brain had demonstrated the production of plasminogen activator (PA) by developing mouse cerebellum. This thesis describes neuronal PA as being primarily tissue plasminogen activator (tPA). This tPA was isolated from a mouse neuronal cell line, and a highly specific antibody was developed in rabbits. The antibody isolated from rabbit antiserum effectively and specifically inhibits murine tPA and blocks the catalytic activity of both the intact molecule and its B-chain, while exhibiting little or nor reactivity with mouse or human urokinase (uPA), human plasmin or plasminogen. This antibody was used as an immunoaffinity ligand to obtain highly purified murine tPA to investigate further the interactions of the protein with cerebellar neurons. Using {sup 125}I-tPA, the high-affinity binding to cerebellar granule neurons is rapid, time-dependent, saturable, reversible and specific for tPA but not for other related serine proteases. Neither the catalytic site nor the carbohydrate moiety of tPA appear to be involved in the binding. Autoradiography shows the specific tPA binding is to granule neurons in these cultures.
- Research Organization:
- Colorado Univ., Denver, CO (USA). Health Sciences Center
- OSTI ID:
- 6709710
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
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PROTEINS
CHEMICAL COMPOSITION
ANTIBODIES
AUTORADIOGRAPHY
BIOCHEMICAL REACTION KINETICS
CEREBELLUM
FRACTIONATION
IODINE 125
LIGANDS
MICE
NERVE CELLS
PLASMINOGEN
TIME DEPENDENCE
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BODY
BRAIN
CENTRAL NERVOUS SYSTEM
DAYS LIVING RADIOISOTOPES
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
FIBRINOLYTIC AGENTS
HEMATOLOGIC AGENTS
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPES
KINETICS
MAMMALS
NERVOUS SYSTEM
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
RADIOISOTOPES
REACTION KINETICS
RODENTS
SEPARATION PROCESSES
SOMATIC CELLS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques