A novel soluble protein factor with non-opioid dynorphin A-binding activity.
A novel soluble non-opioid dynorphin A-binding factor (DABF) was identified and characterized in neuronal cell lines, rat spinal cord, and brain. DABF binds dynorphin A(1-17), dynorphin A(2-17), and the 32 amino acid prodynorphin fragment big dynorphin consisting of dynorphin A and B, but not other opioid and non-opioid peptides, opiates, and benzomorphans. The IC50 for dynorphin A(1-17), dynorphin A(2-17), and big dynorphin is in the 5-10 nM range. Using dynorphin A and big dynorphin fragments a binding epitope was mapped to dynorphin A(6-13). DABF has a molecular mass of about 70 kDa. SH-groups are apparently involved in the binding of dynorphin A since p-hydroxy-mercuribenzoic acid inhibited this process. Upon interaction with DABF dynorphin A was converted into Leu-enkephalin, which remained bound to the protein. These data suggest that DABF functions as an oligopeptidase that forms stable and specific complexes with dynorphin A. The presence of DABF in brain structures and other tissues with low level of prodynorphin expression suggests that DABF as an oligopeptidase may degrade other peptides. Dynorphin A at the sites of its release in the CNS may attenuate this degradation as a competitor when it specifically binds to the enzyme.
Marinova
Zoya
Z
Alcohol and Drug Dependence Research Section, Department of Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden.
Yakovleva
Tatjana
T
Melzig
Matthias F
MF
Hallberg
Mathias
M
Nylander
Ingrid
I
Ray
Kallol
K
Rodgers
David W
DW
Hauser
Kurt F
KF
Ekström
Tomas J
TJ
Bakalkin
Georgy
G
D000818:Animals; D001921:Brain / Q000378:metabolism; D018288:Carcinoma, Small Cell; D002352:Carrier Proteins / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D002822:Choriocarcinoma; D003634:DDT / Q000494:pharmacology; D004399:Dynorphins / Q000378:metabolism*; D006801:Humans; D006902:Hydroxymercuribenzoates / Q000494:pharmacology; D007700:Kinetics; D008175:Lung Neoplasms; D051379:Mice; D008815:Mice, Inbred Strains; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009447:Neuroblastoma; D009474:Neurons / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology
D002352:Carrier Proteins; D006902:Hydroxymercuribenzoates; D009419:Nerve Tissue Proteins; D011480:Protease Inhibitors; C030601:4-hydroxymercuribenzoate; D004399:Dynorphins; D003634:DDT