TY - JOUR
T1 - Molecular and Physiological Characterization of a Receptor for d -Amino Acid-Containing Neuropeptides
AU - Checco, James W.
AU - Zhang, Guo
AU - Yuan, Wang Ding
AU - Yu, Ke
AU - Yin, Si Yuan
AU - Roberts-Galbraith, Rachel H.
AU - Yau, Peter M.
AU - Romanova, Elena V.
AU - Jing, Jian
AU - Sweedler, Jonathan V.
N1 - Funding Information:
The authors thank P. Bauknecht and G. Jeḱ ely for the generous gift of plasmids containing apALNR and Gα-16 for mammalian expression and the UIUC Roy J. Carver Biotechnology Center for providing resources, instrument time, and helpful advice. This work was supported by the National Institutes of Health, via Grant P30 DA018310 from the National Institute on Drug Abuse, and Grant R01 NS031609 from the National Institute of Neurological Disorders and Stroke (J.V.S.), the National Resource for Aplysia, funded by PHS grant P40 OD010952, and the National Natural Science Foundation of China, via Grants 31671097, 31371104 (J.J.), and J1103512 and J1210026 (School of Life Sciences). J.W.C. was supported in part by a Beckman Institute Postdoctoral Fellowship, funded by a Beckman Foundation gift to the Beckman Institute for Advanced Science and Technology at the University of Illinois at Urbana-Champaign.
Publisher Copyright:
© 2018 American Chemical Society.
PY - 2018/5/18
Y1 - 2018/5/18
N2 - Neuropeptides in several animals undergo an unusual post-translational modification, the isomerization of an amino acid residue from the l-stereoisomer to the d-stereoisomer. The resulting d-amino acid-containing peptide (DAACP) often displays biological activity higher than that of its all-l-residue analogue, with the d-residue being critical for function in many cases. However, little is known about the full physiological roles played by DAACPs, and few studies have examined the interaction of DAACPs with their cognate receptors. Here, we characterized the signaling of several DAACPs derived from a single neuropeptide prohormone, the Aplysia californica achatin-like neuropeptide precursor (apALNP), at their putative receptor, the achatin-like neuropeptide receptor (apALNR). We first used quantitative polymerase chain reaction and in situ hybridization experiments to demonstrate receptor (apALNR) expression throughout the central nervous system; on the basis of the expression pattern, we identified novel physiological functions that may be mediated by apALNR. To gain insight into ligand signaling through apALNR, we created a library of native and non-native neuropeptide analogues derived from apALNP (the neuropeptide prohormone) and evaluated them for activity in cells co-transfected with apALNR and the promiscuous Gα subunit Gα-16. Several of these neuropeptide analogues were also evaluated for their ability to induce circuit activity in a well-defined neural network associated with feeding behavior in intact ganglia from Aplysia. Our results reveal the specificity of apALNR and provide strong evidence that this receptor mediates diverse physiological functions throughout the central nervous system. Finally, we show that some native apALNP-derived DAACPs exhibit enhanced stability toward endogenous proteases, suggesting that the d-residues in these DAACPs may increase the peptide lifetime, in addition to influencing receptor specificity, in the nervous system. Ultimately, these studies provide insight into signaling at one of the few known DAACP-specific receptors and advance our understanding of the roles that l- to d-residue isomerization play in neuropeptide signaling.
AB - Neuropeptides in several animals undergo an unusual post-translational modification, the isomerization of an amino acid residue from the l-stereoisomer to the d-stereoisomer. The resulting d-amino acid-containing peptide (DAACP) often displays biological activity higher than that of its all-l-residue analogue, with the d-residue being critical for function in many cases. However, little is known about the full physiological roles played by DAACPs, and few studies have examined the interaction of DAACPs with their cognate receptors. Here, we characterized the signaling of several DAACPs derived from a single neuropeptide prohormone, the Aplysia californica achatin-like neuropeptide precursor (apALNP), at their putative receptor, the achatin-like neuropeptide receptor (apALNR). We first used quantitative polymerase chain reaction and in situ hybridization experiments to demonstrate receptor (apALNR) expression throughout the central nervous system; on the basis of the expression pattern, we identified novel physiological functions that may be mediated by apALNR. To gain insight into ligand signaling through apALNR, we created a library of native and non-native neuropeptide analogues derived from apALNP (the neuropeptide prohormone) and evaluated them for activity in cells co-transfected with apALNR and the promiscuous Gα subunit Gα-16. Several of these neuropeptide analogues were also evaluated for their ability to induce circuit activity in a well-defined neural network associated with feeding behavior in intact ganglia from Aplysia. Our results reveal the specificity of apALNR and provide strong evidence that this receptor mediates diverse physiological functions throughout the central nervous system. Finally, we show that some native apALNP-derived DAACPs exhibit enhanced stability toward endogenous proteases, suggesting that the d-residues in these DAACPs may increase the peptide lifetime, in addition to influencing receptor specificity, in the nervous system. Ultimately, these studies provide insight into signaling at one of the few known DAACP-specific receptors and advance our understanding of the roles that l- to d-residue isomerization play in neuropeptide signaling.
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U2 - 10.1021/acschembio.8b00167
DO - 10.1021/acschembio.8b00167
M3 - Article
C2 - 29543428
AN - SCOPUS:85047488457
SN - 1554-8929
VL - 13
SP - 1343
EP - 1352
JO - ACS chemical biology
JF - ACS chemical biology
IS - 5
ER -