Alanine‐(87)‐threonine polymorphism impairs signaling and internalization of the human P2Y11 receptor, when co‐expressed with the P2Y1 receptor. (13th February 2014)

• Record Type: Journal Article
• Authors: Haas, Michael; Shaaban, Ahmed; Reiser, Georg
• Is Part Of: Journal of neurochemistry. Volume 129:Number 4(2014:May)
• Page Start: 602
• Page End: 613
• Publication Date: 2014-02-13
• Subjects: Neurochemistry -- Periodicals; 616.8042
• DOI: 10.1111/jnc.12666
• Languages: English
• ISSNs: 0022-3042

Abstract

<abstract abstract-type="main" id="jnc12666-abs-0001"> <title>Abstract</title> <p>The P2Y<sub>11</sub> nucleotide receptor detects high extracellular ATP concentrations. Mutations of the human <italic>P2RY</italic><sub><italic>11</italic></sub> gene can play a role in brain autoimmune responses, and the P2Y<sub>11</sub> receptor alanine‐87‐threonine (A87T) polymorphism has been suggested to affect immune‐system functions. We investigated receptor functionality of the P2Y<sub>11</sub>A87T mutant using HEK293 and 1321N1 astrocytoma cells. In HEK293 cells, the P2Y<sub>11</sub> receptor agonist 3′‐<italic>O</italic>‐(4‐benzoylbenzoyl)adenosine 5′‐triphosphate (BzATP) was completely inactive in evoking intracellular calcium release while the potency of ATP was reduced. ATP was also less potent in triggering cAMP generation. However, 1321N1 astrocytoma cells, which lack any endogenous P2Y<sub>1</sub> receptors, did not display a reduction. Only when 1321N1 cells were co‐transfected with P2Y<sub>11</sub>A87T and P2Y<sub>1</sub> receptors, the calcium responses to the P2Y<sub>11</sub> receptor‐specific agonist BzATP were reduced. It is already known that P2Y<sub>1</sub> and P2Y<sub>11</sub> receptors interact. We thus conclude that the physiological impact of A87T mutation of the P2Y<sub>11</sub> receptor derives from detrimental effects on P2Y<sub>1</sub>–P2Y<sub>11</sub> receptor interaction. We additionally investigated alanine‐87‐serine and alanine‐87‐tyrosine P2Y<sub>11</sub> receptor mutants. Both mutations rescue the response to BzATP in HEK293 cells, thus ruling out polarity of amino acid‐87 to be the molecular basis for altered receptor characteristics. We further found that the P2Y<sub>11</sub>A87T receptor shows complete loss of nucleotide‐induced internalization in HEK293 cells. Thus, we demonstrate impaired signaling of the P2Y<sub>11</sub> A87T‐mutated receptors when co‐operating with P2Y<sub>1</sub> receptors. <boxed-text content-type="graphic" id="jnc12666-blkfxd-0002" position="anchor" orientation="portrait"><graphic position="anchor" mimetype="image" xlink:href="ark:/27927/pgg5f98pt3p" orientation="portrait" xlink:type="simple" xmlns:xlink="http://www.w3.org/1999/xlink" /></boxed-text></p> <p>The human P2Y<sub>11</sub> nucleotide receptor plays key role in immune‐responses in brain and other tissues. We provide evidence for significant functional disturbance of the P2Y<sub>11</sub> receptor carrying the Alanine‐87‐Threonine mutation caused by natural polymorphism. This receptor defect is apparent only when co‐expressed with P2Y<sub>1</sub> receptors. We found reductions in ligand‐induced calcium and cAMP responses and in nucleotide‐induced receptor internalization / resensitization. Thus, prolonged nucleotide treatments are the basis for the molecular defects of the mutant receptor in diseases.</p> </abstract>