High-performance liquid chromatographic analysis of in vitro central neuropeptide processing.
Reversed-phase high-performance liquid chromatography (HPLC) was used to study and characterize the in vitro proteolytic processing of beta-endorphin by twice-washed membrane homogenates. A high-resolution method, capable of separating over 30 different human beta-endorphin-related fragments in a single analysis, was used to study the time course of production of specific, biologically active endorphin fragments by membrane-associated proteases. The results demonstrate that frozen (-37 degrees C), postmortem human and rat brains are viable for processing studies and that metabolism proceeds similarly to that in fresh brain homogenates or slices. Significant differences were noted in the formation rates of putative neuroleptic peptides between sex- and age-matched postmortem brain tissues from controls versus postmortem brain tissues from neuropsychiatric patients or drug-treated animals. These data suggest that using HPLC to characterize neuropeptide processing in human or rat membrane-associated enzyme homogenates is both descriptive and quantitative and offers insight into the central regulation of neuropeptide metabolism.
T P
TP
Davis
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A
Culling-Berglund
D000595:Amino Acid Sequence; D000818:Animals; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid; D004723:Endorphins / Q000302:isolation & purification; D004798:Enzymes / Q000302:isolation & purification; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008297:Male; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D012997:Solvents; D013045:Species Specificity; D013997:Time Factors; D018822:alpha-Endorphin; D001615:beta-Endorphin; D018823:gamma-Endorphin
D004723:Endorphins; D004798:Enzymes; D009419:Nerve Tissue Proteins; D012997:Solvents; D001615:beta-Endorphin; D018822:alpha-Endorphin; D018823:gamma-Endorphin