dc.contributor.author | Zhang, J | |
dc.contributor.author | Siew, K | |
dc.contributor.author | Macartney, T | |
dc.contributor.author | O'Shaughnessy, KM | |
dc.contributor.author | Alessi, DR | |
dc.date.accessioned | 2018-04-04T10:14:35Z | |
dc.date.accessioned | 2018-07-09T14:19:02Z | |
dc.date.issued | 2015-05-20 | |
dc.description.abstract | The STE20/SPS1-related proline/alanine-rich kinase (SPAK) controls blood pressure (BP) by phosphorylating and stimulating the Na-Cl (NCC) and Na-K-2Cl (NKCC2) co-transporters, which regulate salt reabsorption in the kidney. SPAK possesses a conserved carboxy-terminal (CCT) domain, which recognises RFXV/I motifs present in its upstream activator [isoforms of the With-No-lysine (K) kinases (WNKs)] as well as its substrates (NCC and NKCC2). To define the physiological importance of the CCT domain, we generated knock-in mice in which the critical CCT domain Leu502 residue required for high affinity recognition of the RFXI/V motif was mutated to Alanine. The SPAK CCT domain defective knock-in animals are viable, and the Leu502Ala mutation abolished co-immunoprecipitation of SPAK with WNK1, NCC and NKCC2. The CCT domain defective animals displayed markedly reduced SPAK activity and phosphorylation of NCC and NKCC2 co-transporters at the residues phosphorylated by SPAK. This was also accompanied by a reduction in the expression of NCC and NKCC2 protein without changes in mRNA levels. The SPAK CCT domain knock-in mice showed typical features of Gitelman Syndrome with mild hypokalaemia, hypomagnesaemia, hypocalciuria and displayed salt wasting on switching to a low-Na diet. These observations establish that the CCT domain plays a crucial role in controlling SPAK activity and BP. Our results indicate that CCT domain inhibitors would be effective at reducing BP by lowering phosphorylation as well as expression of NCC and NKCC2. | en_GB |
dc.description.sponsorship | This work was supported by the Medical Research Council, British Heart foundation (a PhD studentship to K.S. and PG 13 89 30577), and the pharmaceutical companies supporting the Division of Signal Transduction Therapy Unit (AstraZeneca, Boehringer-Ingelheim, GlaxoSmithKline, Merck KGaA, Janssen Pharmaceutica and Pfizer). Funding to pay the Open Access publication charges for this article was provided by the University of Cambridge RCUK and COAF block grants. | en_GB |
dc.identifier.citation | Vol. 24 (16), pp. 4545 - 4558 | en_GB |
dc.identifier.doi | 10.1093/hmg/ddv185 | |
dc.identifier.uri | http://hdl.handle.net/10871/33414 | |
dc.language.iso | en | en_GB |
dc.publisher | Oxford University Press (OUP) | en_GB |
dc.relation.url | https://www.ncbi.nlm.nih.gov/pubmed/25994507 | en_GB |
dc.relation.url | http://hdl.handle.net/10871/32308 | en_GB |
dc.rights | © The Author 2015. Published by Oxford University Press.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. | en_GB |
dc.subject | Amino Acid Substitution | en_GB |
dc.subject | Animals | en_GB |
dc.subject | Blood Pressure | en_GB |
dc.subject | Gitelman Syndrome | en_GB |
dc.subject | HEK293 Cells | en_GB |
dc.subject | Humans | en_GB |
dc.subject | Mice | en_GB |
dc.subject | Mice, Mutant Strains | en_GB |
dc.subject | Mutation, Missense | en_GB |
dc.subject | Phosphorylation | en_GB |
dc.subject | Protein Structure, Tertiary | en_GB |
dc.subject | Protein-Serine-Threonine Kinases | en_GB |
dc.subject | Solute Carrier Family 12, Member 3 | en_GB |
dc.title | Critical role of the SPAK protein kinase CCT domain in controlling blood pressure | en_GB |
dc.type | Article | en_GB |
dc.date.available | 2018-04-04T10:14:35Z | |
dc.date.available | 2018-07-09T14:19:02Z | |
exeter.place-of-publication | England | en_GB |
dc.description | This is the final version of the article. Available from OUP via the DOI in this record. | en_GB |
dc.description | There is another ORE record for this publication: http://hdl.handle.net/10871/32308 | en_GB |
dc.identifier.journal | Human Molecular Genetics | en_GB |