Combined antibody-high-performance liquid chromatography approach to assess prohormone processing

Publisher Summary This chapter discusses the combined antibody-high-performance liquid chromatography approach to assess prohormone processing. This approach creates special problems for the protein biochemist and physiologist because the amino acid sequences of the various peptides may not be known with certainty (the peptide has not been sequenced, only deduced from a complementary DNA (cDNA)), the regulatory steps involved in the biosynthesis, processing, and secretion of the peptides are unclear, and their functional roles are often unexplored. Various tissues may metabolize the same precursor peptides by different routes, and these products may have different functional consequences depending on the locus of secretion and the physiological/pathological status of the organism. These circumstances emphasize the importance of adopting a coordinated and multifaceted approach in detailing a peptide's fate from biosynthesis to function. Perhaps, through additional biochemical and physiological manipulations, we will be able to dissect more clearly the contribution of different gonadal factors to the biosynthesis, processing, transport, secretion, and/or degradation of these various processed products from luteinizing hormone releasing hormone (LHRH) neurons.