Inositol 1,4,5-trisphosphate (IP3) receptors from cerebellum and recombinant type 1 IP3 receptors expressed in Sf9 cells had indistinguishable affinities for IP3 (Kd = 6.40±0.48nM) and adenophostin A (Kd = 0.89±0.05nM). In cytosol-like medium, each of the three mammalian IP3 receptor subtypes when expressed in Sf9 cells bound adenophostin A with greater affinity than IP3. It has been suggested that adenophostin A binds with high affinity only in the presence of ATP, but we found that adenophostin A similarly displaced [3H]IP3 from type 1 IP3 receptors whatever the ATP concentration. N-terminal fragments of the type 1 receptor were expressed with and without the S1 splice site; its removal had no effect on [3H]IP3 binding to the 1—604 protein, but abolished binding to the 224—604 protein. The 1—604 fragment and full-length receptor bound adenophostin A with the same affinity, but the fragment had 3-fold greater affinity for IP3, suggesting that C-terminal residues selectively inhibit IP3 binding. The 224—604S1+ fragment bound IP3 and adenophostin A with increased affinity, but as with the 1—604 fragment it bound adenophostin A with only 2-fold greater affinity than IP3. High-affinity binding of adenophostin A may be partially determined by its 2′-phosphate interacting more effectively than the 1-phosphate of IP3 with residues within the IP3-binding core. This may account for the 2-fold greater affinity of adenophostin A relative to IP3 for the minimal IP3-binding domain. In addition we suggest that C-terminal residues, which impede access of IP3, may selectively interact with adenophostin A to allow it unhindered access to the IP3-binding domain.
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October 2002
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Research Article|
October 01 2002
Determinants of adenophostin A binding to inositol trisphosphate receptors Available to Purchase
Stephen A. MORRIS;
Stephen A. MORRIS
∗Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1PD, U.K.,
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Edmund P. NEROU;
Edmund P. NEROU
∗Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1PD, U.K.,
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Andrew M. RILEY;
Andrew M. RILEY
†Wolfson Laboratory of Medicinal Chemistry, Department of Pharmacy and Pharmacology, University of Bath, Claverton Down, Bath, BA2 7AY, U.K.
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Barry V.L. POTTER;
Barry V.L. POTTER
†Wolfson Laboratory of Medicinal Chemistry, Department of Pharmacy and Pharmacology, University of Bath, Claverton Down, Bath, BA2 7AY, U.K.
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Colin W. TAYLOR
Colin W. TAYLOR
1
∗Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1PD, U.K.,
1To whom correspondence should be addressed (e-mail [email protected]).
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Publisher: Portland Press Ltd
Received:
April 29 2002
Revision Received:
June 25 2002
Accepted:
June 27 2002
Accepted Manuscript online:
June 27 2002
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2002
2002
Biochem J (2002) 367 (1): 113–120.
Article history
Received:
April 29 2002
Revision Received:
June 25 2002
Accepted:
June 27 2002
Accepted Manuscript online:
June 27 2002
Citation
Stephen A. MORRIS, Edmund P. NEROU, Andrew M. RILEY, Barry V.L. POTTER, Colin W. TAYLOR; Determinants of adenophostin A binding to inositol trisphosphate receptors. Biochem J 1 October 2002; 367 (1): 113–120. doi: https://doi.org/10.1042/bj20020675
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