Enkephalin-containing polypeptides in human cerebrospinal fluid.
The chemical nature of peptides in human CSF with the enkephalin core sequence from proenkephalin A and proenkephalin B, was investigated. Direct measurements with radioimmunoassay (RIA) were run on enkephalin, enkephalin hexapeptides, dynorphin A, dynorphin A1-8 and dynorphin B. The hexapeptides occurred in about 3 times higher concentration than the corresponding enkephalins. The only dynorphin RIA which gave positive results was the one for dynorphin A. However, most dynorphin A immunoactive material showed higher apparent molecular weight (MW; 3 and 5 kdalton) than the standard (2 kdalton). To identify and quantitate every possible proenkephalin derived peptide with the enkephalin sequence, chromatographic fractions were treated with trypsin. The products, Leu-enkephalin-Arg6 (from proenkephalin B) and Met-enkephalin-Lys6/Arg6 (from proenkephalin A) were measured by specific RIAs and identified by HPLC. In the higher (greater than 5 kdalton) MW interval, there was about 10-fold higher yield of Met-enkephalyl than Leu-enkephalyl hexapeptides. In the intermediate 1-3 kdalton MW interval, most activity derived from proenkephalin B. Finally, from the low MW region, there was about 5 times more proenkephalin A peptides. The main dynorphin A peak at 5 kdalton was transferred to a major Leu-enkephalin-Arg6 peak by trypsin degradation. The data indicate the presence of a whole family of peptides from the two proenkephalin genes in human CSF. Precursors to the peptides supposed to be the active members in the proenkephalin families occur in relatively high concentrations and may provide good markers for activity in these peptide systems.
Nyberg
F
F
Nylander
I
I
Terenius
L
L
D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004399:Dynorphins / Q000134:cerebrospinal fluid / Q000302:isolation & purification; D004743:Enkephalin, Leucine / Q000134:cerebrospinal fluid / Q000302:isolation & purification; D004744:Enkephalin, Methionine / Q000134:cerebrospinal fluid / Q000302:isolation & purification; D004745:Enkephalins / Q000134:cerebrospinal fluid* / Q000302:isolation & purification; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D010455:Peptides / Q000134:cerebrospinal fluid* / Q000302:isolation & purification; D011498:Protein Precursors / Q000134:cerebrospinal fluid* / Q000302:isolation & purification; D011863:Radioimmunoassay
D004745:Enkephalins; D010455:Peptides; D011498:Protein Precursors; C029992:proenkephalin; D004744:Enkephalin, Methionine; D004743:Enkephalin, Leucine; D004399:Dynorphins