Evidence for the abundant expression of arginine 185 containing human CRF2α receptors and the role of position 185 for receptor-ligand selectivity

Urocortin 2 treatment is protective in excitotoxic retinal degeneration

“…The receptor subtype underlying the presently shown retinoprotective action of Ucn 2 remains to be determined. In the retina the two CRF receptor subtypes are present at both mRNA and protein levels (10,40). Based on the present findings, the relevance of endogenous CRF/Ucns to retinal function should be further explored.…”

Urocortin 2 treatment is protective in excitotoxic retinal degeneration

“…The receptor subtype underlying the presently shown retinoprotective action of Ucn 2 remains to be determined. In the retina the two CRF receptor subtypes are present at both mRNA and protein levels (10,40). Based on the present findings, the relevance of endogenous CRF/Ucns to retinal function should be further explored.…”

Structural and Functional Insights into CRF Peptides and Their Receptors

“…A different study has also shown that position 185 EL1 of human CRF 2 R (189 EL1 of human CRF 1 R) plays a role in receptor function. Specifically, SVG and UI bound with increased affinities to CRF 2 R when arginine was at position 185 EL1 compared to the presence of histidine at this position [107]. In addition, the substitution of Tyr346 EL3 , Phe347 EL3, and Asn348 EL3 of CRF 1 R by Ala significantly reduced the binding affinity of CRF [108].…”

The highly selective CRF2 receptor antagonist K41498 binds to presynaptic CRF2 receptors in rat brain

“…Binding of the aSvg‐30 analogue to the hCRF 1 , hCRF 2α or hCRF 2β receptor was performed essentially as described previously (Dautzenberg et al ., 2000, 2001b). Briefly, membranes isolated from HEK293 cells stably expressing the hCRF 1 (30 μg), hCRF 2α (5 μg) or hCRF 2β (5 μg) receptor were combined with 100 p M 125 I‐Tyr 0 ‐Svg (Amersham, Little Chalfont, U.K., 2000 Ci/mmol).…”

“…Briefly, membranes isolated from HEK293 cells stably expressing the hCRF 1 (30 μg), hCRF 2α (5 μg) or hCRF 2β (5 μg) receptor were combined with 100 p M 125 I‐Tyr 0 ‐Svg (Amersham, Little Chalfont, U.K., 2000 Ci/mmol). Binding was performed in 96‐well plates (Beckmann Instruments, Fullertown, CA, U.S.A.) with 0.5 mg wheat germ agglutinin beads using a scintillation proximity assay as described previously (Dautzenberg et al ., 2000; Higelin et al ., 2001). The inhibition constant ( K i ) was calculated using the interactive curve‐fitting program Xlfit (Dautzenberg et al ., 2001b).…”

Urocortin 2 treatment is protective in excitotoxic retinal degeneration

“…The receptor subtype underlying the presently shown retinoprotective action of Ucn 2 remains to be determined. In the retina the two CRF receptor subtypes are present at both mRNA and protein levels (10,40). Based on the present findings, the relevance of endogenous CRF/Ucns to retinal function should be further explored.…”

Structural and Functional Insights into CRF Peptides and Their Receptors

“…A different study has also shown that position 185 EL1 of human CRF 2 R (189 EL1 of human CRF 1 R) plays a role in receptor function. Specifically, SVG and UI bound with increased affinities to CRF 2 R when arginine was at position 185 EL1 compared to the presence of histidine at this position [107]. In addition, the substitution of Tyr346 EL3 , Phe347 EL3, and Asn348 EL3 of CRF 1 R by Ala significantly reduced the binding affinity of CRF [108].…”

The highly selective CRF2 receptor antagonist K41498 binds to presynaptic CRF2 receptors in rat brain

“…Binding of the aSvg‐30 analogue to the hCRF 1 , hCRF 2α or hCRF 2β receptor was performed essentially as described previously (Dautzenberg et al ., 2000, 2001b). Briefly, membranes isolated from HEK293 cells stably expressing the hCRF 1 (30 μg), hCRF 2α (5 μg) or hCRF 2β (5 μg) receptor were combined with 100 p M 125 I‐Tyr 0 ‐Svg (Amersham, Little Chalfont, U.K., 2000 Ci/mmol).…”

“…Briefly, membranes isolated from HEK293 cells stably expressing the hCRF 1 (30 μg), hCRF 2α (5 μg) or hCRF 2β (5 μg) receptor were combined with 100 p M 125 I‐Tyr 0 ‐Svg (Amersham, Little Chalfont, U.K., 2000 Ci/mmol). Binding was performed in 96‐well plates (Beckmann Instruments, Fullertown, CA, U.S.A.) with 0.5 mg wheat germ agglutinin beads using a scintillation proximity assay as described previously (Dautzenberg et al ., 2000; Higelin et al ., 2001). The inhibition constant ( K i ) was calculated using the interactive curve‐fitting program Xlfit (Dautzenberg et al ., 2001b).…”

Urocortin 2 treatment is protective in excitotoxic retinal degeneration

“…The receptor subtype underlying the presently shown retinoprotective action of Ucn 2 remains to be determined. In the retina the two CRF receptor subtypes are present at both mRNA and protein levels (10,40). Based on the present findings, the relevance of endogenous CRF/Ucns to retinal function should be further explored.…”

Structural and Functional Insights into CRF Peptides and Their Receptors

“…A different study has also shown that position 185 EL1 of human CRF 2 R (189 EL1 of human CRF 1 R) plays a role in receptor function. Specifically, SVG and UI bound with increased affinities to CRF 2 R when arginine was at position 185 EL1 compared to the presence of histidine at this position [107]. In addition, the substitution of Tyr346 EL3 , Phe347 EL3, and Asn348 EL3 of CRF 1 R by Ala significantly reduced the binding affinity of CRF [108].…”

The highly selective CRF2 receptor antagonist K41498 binds to presynaptic CRF2 receptors in rat brain

“…Binding of the aSvg‐30 analogue to the hCRF 1 , hCRF 2α or hCRF 2β receptor was performed essentially as described previously (Dautzenberg et al ., 2000, 2001b). Briefly, membranes isolated from HEK293 cells stably expressing the hCRF 1 (30 μg), hCRF 2α (5 μg) or hCRF 2β (5 μg) receptor were combined with 100 p M 125 I‐Tyr 0 ‐Svg (Amersham, Little Chalfont, U.K., 2000 Ci/mmol).…”

“…Briefly, membranes isolated from HEK293 cells stably expressing the hCRF 1 (30 μg), hCRF 2α (5 μg) or hCRF 2β (5 μg) receptor were combined with 100 p M 125 I‐Tyr 0 ‐Svg (Amersham, Little Chalfont, U.K., 2000 Ci/mmol). Binding was performed in 96‐well plates (Beckmann Instruments, Fullertown, CA, U.S.A.) with 0.5 mg wheat germ agglutinin beads using a scintillation proximity assay as described previously (Dautzenberg et al ., 2000; Higelin et al ., 2001). The inhibition constant ( K i ) was calculated using the interactive curve‐fitting program Xlfit (Dautzenberg et al ., 2001b).…”