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
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ANIMALS
ANTIBODIES
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BODY
BRAIN
CENTRAL NERVOUS SYSTEM
CEREBELLUM
CHEMICAL COMPOSITION
DAYS LIVING RADIOISOTOPES
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
FIBRINOLYTIC AGENTS
FRACTIONATION
HEMATOLOGIC AGENTS
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPES
KINETICS
LIGANDS
MAMMALS
MICE
NERVE CELLS
NERVOUS SYSTEM
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
PLASMINOGEN
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RODENTS
SEPARATION PROCESSES
SOMATIC CELLS
TIME DEPENDENCE
VERTEBRATES