SUMMARYThe effects of several N-sulfonyl-polyamines, including N 1 -dansyl-spermine (N 1 -DnsSpm) and N 1 -(n-octanesulfonyl)-spermine (N 1 -OsSpm), were studied at recombinant N-methyl-D-aspartate (NMDA) receptors expressed in Xenopus laevis oocytes. N 1 -DnsSpm and N 1 -OsSpm inhibited NMDA receptors and were ϳ1000-fold more potent than spermine in oocytes voltage-clamped at Ϫ70 mV. Block by N 1 -DnsSpm and N 1 -OsSpm was strongly voltage dependent, being more pronounced at hyperpolarized membrane potentials. With the Woodhull model of voltage-dependent channel block, the values of K d (0) were 779 M, 882 M, and 7.4 mM and those of z␦ were 2.58, 2.57, and 1.07 for N 1 -DnsSpm, N 1 -OsSpm, and spermine, respectively. This suggests that an increase in the voltage dependence of block together with an increase in affinity contributes to the increased potencies of N 1 -DnsSpm and N 1 -OsSpm compared with spermine. Sensitivity to N 1 -DnsSpm was reduced by mutation NR1(N616Q) and was increased by mutations NR1(N616G) and NR2A(N615G). The NR1(N616G) and NR2A(N615G) mutations decreased the K d (0) value of N 1 -DnsSpm without affecting z␦, whereas the NR1(N616Q) mutation reduced z␦. These mutations may alter the accessibility of part of the polyamine binding site within the channel pore or directly alter the properties of that site. Block by N 1 -DnsSpm (0.3 M) was almost complete at Ϫ100 mV, and there was no relief of block at extreme negative membrane potentials (Ϫ100 to Ϫ200 mV) at wild-type NR1/NR2A channels. In contrast, block by N 1 -DnsSpm was partially relieved at extreme negative potentials at receptors containing NR1(N616G) or NR2A(N615G), suggesting that N 1 -DnsSpm can permeate these mutant channels but not wild-type NR1/NR2A channels. This is hypothesized to be due to an increase in the pore size of channels containing NR1(N616G) or NR2A(N615G), which allows passage of the bulky head group of N 1 -DnsSpm. In contrast to N 1 -DnsSpm, N 1 -OsSpm could easily permeate wildtype NR1/NR2A channels, presumably because the head group of N 1 -OsSpm can pass through the narrowest part of the channel pore. N-Sulfonyl-polyamines such as N 1 -DnsSpm and N 1 -OsSpm represent a new class of polyamine antagonists with which to study glutamate receptor ion channels.The endogenous polyamine spermine has a variety of effects on NMDA and non-NMDA glutamate receptors (1, 2). At NMDA receptors, spermine has both stimulatory and inhibitory effects when applied extracellularly (3-7). Inhibition of NMDA receptors by spermine is strongly voltage dependent and may be caused by an open-channel block and/or screening of surface charges around the mouth or vestibule of the ion channel (3,5,8). Intracellular spermine can block the ion channel of some subtypes of AMPA and kainate receptors, an effect that is responsible for inward rectification of these receptors (9 -12) and may be mechanistically similar to the block of inward-rectifier K ϩ channels by polyamines (13,14). When applied extracellularly, spermine is a relatively weak anta...