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J. Gen. Physiol.,
Volume 110, Number 5, November 1, 1997 485-502
From the Department of Biophysical Sciences, State University of New York, Buffalo, New York 14214
Single-channel currents were recorded from mouse NR1-NR2B (
-
2) receptors containing mixtures
of wild-type and mutant subunits expressed in Xenopus oocytes. Mutant subunits had an asparagine-to-glutamine (N-to-Q) mutation at the N0 site of the M2 segment (NR1:598, NR2B:589). Receptors with pure N or Q NR1 and
NR2 subunits generated single-channel currents with distinctive current patterns. Based on main and sublevel amplitudes, occupancy probabilities, and lifetimes, four patterns of current were identified, corresponding to receptors with the following subunit compositions (NR1/NR2): N/N, N/Q, Q/N, and Q/Q. Only one current pattern
was apparent for each composition. When a mixture of N and Q NR2 subunits was coexpressed with pure mutant
NR1 subunits, three single-channel current patterns were apparent. One pattern was the same as Q/Q receptors
and another was the same as Q/N receptors. The third, novel pattern presumably arose from hybrid receptors
having both N and Q NR2 subunits. When a mixture of N and Q NR1 subunits was coexpressed with pure mutant
NR2 subunits, six single-channel current patterns were apparent. One pattern was the same as Q/Q receptors and
another was the same as N/Q receptors. The four novel patterns presumably arose from hybrid receptors having
both N and Q NR1 subunits. The relative frequency of NR1 hybrid receptor current patterns depended on the relative amounts of Q and N subunits that were injected into the oocytes. The number of hybrid receptor patterns suggests that there are two NR2 subunits per receptor and is consistent with either three or five NR1 subunits per
receptor, depending on whether or not the order of mutant and wild-type subunits influences the current pattern.
When considered in relation to other studies, the most straightforward interpretation of the results is that N-methyl-D-aspartate receptors are pentamers composed of three NR1 and two NR2 subunits.
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