Table 1. K d values determined for GluA2... | Rockefeller University Press

Table 1.

K_d values determined for GluA2 constructs by different techniques

Technique	Temperature	GluA2S	GnTI⁻	EndoH	GluA2L	GluA2S-FAM	EndoH-FAM	EndoH-DyLight
SV	10°C	6.0 [2.1–22]^a				14.9^b [7.6–47]
	20°C	8.3 [2.0–22]	11 [0.8–43]	5.6 [2.3–23]	5.5 [2.8–43]
				8.3 [5.1–27]
				9.7 [4.4–16]
FDS-SV	20°C						5.3^c [3.0–14]
							26^d [10–57]
SE	4°C			160 [83–268]
				284 [159–461]
				30^e [ND–260]
	10°C			13^f [0.14–50]
				17 [ND–74]
				244^f [46–643]
FAI	20°C							10.8 [2.4–29]
								11.3 [ND–68]
								8.43 [ND–50.8]

Technique	Temperature	GluA2S	GnTI⁻	EndoH	GluA2L	GluA2S-FAM	EndoH-FAM	EndoH-DyLight
SV	10°C	6.0 [2.1–22]^a				14.9^b [7.6–47]
	20°C	8.3 [2.0–22]	11 [0.8–43]	5.6 [2.3–23]	5.5 [2.8–43]
				8.3 [5.1–27]
				9.7 [4.4–16]
FDS-SV	20°C						5.3^c [3.0–14]
							26^d [10–57]
SE	4°C			160 [83–268]
				284 [159–461]
				30^e [ND–260]
	10°C			13^f [0.14–50]
				17 [ND–74]
				244^f [46–643]
FAI	20°C							10.8 [2.4–29]
								11.3 [ND–68]
								8.43 [ND–50.8]

K_d values for individual experiments are reported in nanomolar; errors represent the 95% confidence interval using an automated surface projection method, unless indicated otherwise. 10 independent AUC experiments were performed for GluA2S digested with EndoH, as indicated by replicate entries for the mean and 95% CI. SV, sedimentation velocity with absorbance and interference optics; FDS-SV, sedimentation velocity with fluorescence detection optics; SE, sedimentation equilibrium with absorbance optics; FAI, fluorescence anisotropy.

68.3% confidence interval.

The data obtained after FAM labeling of GluA2 led to the highest best-fit value, but the labeling does not significantly affect binding within the 95% confidence interval of this assay.

Analysis of FDS-SV data only.

Analysis of FDS data with single high concentration data point measured by absorbance at 488 nm.

Analysis of data with low loading concentration and 210-nm detection.

Analysis of data with oil layer to increase pressure.

[ViewLarge]