HUMAN LIPOXYGENASE PRUFICATION AND MOLECULAR PROBES

LIPOXGENASES CONSTITUTE A FAMLY OF NON-HEME IRON-CONTAINING DIOXYGENASES THAT HAVE BEEN DEMONSTRATED TO PRODUCE A WIDE RANGE PHYSIOLOGICALLY ACTIVE METABOITES. THE ENZYME 12-LIPOXYGENASE (12-LX) IS PRESENT IN PLATELETS, LEUKOSYTES, LUNG AND BRAIN. 12-LX CONVERTS ARACHIDONIC ACID TO 12-HPETE, WHICH CAN THEN BE CONVERTED TO 12-HETE. MORE RECENTLY, EPOXY (HEPOXYLINS A3 AND B3) DIHYDROXY AND TRIHYDROXY DERIVATIVES OF 12(S)-HPETE HAVE BEEN DESCRIBED. THE ENZYME 5-LIPOXYGENASE (5-LX) PLAYS A PIVOTAL ROLE IN THE BIOSYNTHESIS OF LEUKOTRIENES, WHICH ARE IMPORTANT IN THE REGULATION OF NUMEROUS BIOLOGICAL PROCESSES, RANGING FROM ACUTE INFLAMMATION AND THROMBOSIS TO CHRONIC CARDIOVASCULAR DISORDERS SUCH AS ATHEROSCLEROSIS. THE ENZYME 15-LIPOXYGENASE (15-LX) PEROXIDIZES ARACHIDONIC ACID TO FORM 15(S)-HPETE, WHICH CAN BE CONVERTED TO 15(S)-HETE AND THE TRIHYDROXY COMPOUNDS LIPOXIN A4 AND LIPOXIN B4, WHICH EXHIBIT A RANGE OF BIOLOGICAL ACTIVITIES. THE ULTIMATE GOAL OF THIS PROJECT IS THE DEVELOPMENT OF COMMERCIALLY VIABLE METHODS FOR PURIFICATION TO HOMOGENEITY OF HUMAN LLIPOXYGENASES,THE PRODUCTION OF POLYCLONAL AND MONOCLONAL ANTIBODIES, AS WELL AS NUCLEIC ACID PROBES SPECIFIC FOR EACH OF THESE ENZYMES. THESE REAGENTS WILL BE MARKETED TO SCIENTISTS INVESTIGATING THE DISTRIBUTION AND REGULATION OF HUMAN 5- AND 15-LIPOXYGENASE ENZYMES IN NORMAL AND DISEASED INDIVIDUALS.