Peptide binding and NMR analysis of the interaction between SAP97 PDZ2 and GluR-A: potential involvement of a disulfide bond

Biochemistry
Lotta von OssowskiKari Keinänen

Abstract

Synaptic delivery of GluR-A (GluR1) subunit-containing glutamate receptors depends on a C-terminal type I PDZ binding motif in GluR-A. Synapse-associated protein 97 (SAP97) is the only PDZ domain protein known to associate with GluR-A. We have used NMR spectroscopy and a biotinylated peptide binding assay to characterize the interaction between synthetic GluR-A C-terminal peptides and the PDZ2 domain of SAP97 (SAP97(PDZ2)), previously determined to be the dominant factor responsible for the interaction. The binding mode appeared to be strongly influenced by redox conditions. Chemical shift changes observed in NMR spectra indicate that under reducing conditions, the last four residues of GluR-A peptides bind to PDZ2 in a fashion typical of class I PDZ interactions. The binding is weak and relatively nonselective as it occurs similarly with a PDZ2 domain derived from PSD-95, a related protein not believed to directly interact with GluR-A. In the absence of reducing agents, conserved cysteine residues in SAP97(PDZ2) and the GluR-A C-terminus gave rise to an anomalous behavior in a microplate assay with a biotinylated GluR-A 18-mer peptide. A covalent disulfide-linked complex between SAP97(PDZ2) and the GluR-A peptide was seen in t...Continue Reading

References

Jun 20, 1997·Journal of Molecular Biology·D E ZimmermanG T Montelione
Oct 13, 2000·The European Journal of Neuroscience·J G ValtschanoffR J Weinberg
Jun 8, 2002·Annual Review of Neuroscience·Roberto Malinow, Robert C Malenka
Oct 1, 2004·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Torsten W SchulzPavel Osten

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Citations

May 13, 2010·Journal of Molecular Modeling·Kristian KaufmannJens Meiler
Sep 16, 2008·Journal of the American Society for Mass Spectrometry·Liwen ZhangYeong-Renn Chen
Oct 10, 2009·Journal of the American Society for Mass Spectrometry·Hongmei YangShuying Liu
Sep 3, 2010·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Mike T LinJohn P Adelman
Oct 31, 2006·The FEBS Journal·Ingemar von OssowskiKari Keinänen
Feb 6, 2017·PloS One·Lotta von OssowskiKari Keinänen
Jun 25, 2009·Analytical Chemistry·Sheng Yu HuangYueh Yi Lin
Dec 31, 2021·Cellular and Molecular Life Sciences : CMLS·Paul BaudinVincent Navarro

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