Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
J Biol Chem
2014 Oct 17;28942:29273-84. doi: 10.1074/jbc.M114.604009.
Show Gene links
Show Anatomy links
Short forms of Ste20-related proline/alanine-rich kinase (SPAK) in the kidney are created by aspartyl aminopeptidase (Dnpep)-mediated proteolytic cleavage.
Markadieu N
,
Rios K
,
Spiller BW
,
McDonald WH
,
Welling PA
,
Delpire E
.
???displayArticle.abstract???
The Ste20-related kinase SPAK regulates sodium, potassium, and chloride transport in a variety of tissues. Recently, SPAK fragments, which lack the catalytic domain and are inhibitory to Na(+) transporters, have been detected in kidney. It has been hypothesized that the fragments originate from alternative translation start sites, but their precise origin is unknown. Here, we demonstrate that kidney lysate possesses proteolytic cleavage activity toward SPAK. Ion exchange and size exclusion chromatography combined with mass spectrometry identified the protease as aspartyl aminopeptidase. The presence of the protease was verified in the active fractions, and recombinant aspartyl aminopeptidase recapitulated the cleavage pattern observed with kidney lysate. Identification of the sites of cleavage by mass spectrometry allowed us to test the function of the smaller fragments and demonstrate their inhibitory action toward the Na(+)-K(+)-2Cl(-) cotransporter, NKCC2.
Bylander,
Human and mouse homo-oligomeric meprin A metalloendopeptidase: substrate and inhibitor specificities.
2007, Pubmed
Bylander,
Human and mouse homo-oligomeric meprin A metalloendopeptidase: substrate and inhibitor specificities.
2007,
Pubmed
CLELAND,
DITHIOTHREITOL, A NEW PROTECTIVE REAGENT FOR SH GROUPS.
1964,
Pubmed
Chaikuad,
Structure of human aspartyl aminopeptidase complexed with substrate analogue: insight into catalytic mechanism, substrate specificity and M18 peptidase family.
2012,
Pubmed
Chen,
Insights into substrate specificity and metal activation of mammalian tetrahedral aspartyl aminopeptidase.
2012,
Pubmed
Chow,
Mammalian pitrilysin: substrate specificity and mitochondrial targeting.
2009,
Pubmed
Correa,
A role for a TIMP-3-sensitive, Zn(2+)-dependent metalloprotease in mammalian gamete membrane fusion.
2000,
Pubmed
Cousins,
Integrative aspects of zinc transporters.
2000,
Pubmed
Delpire,
SPAK and OSR1: STE20 kinases involved in the regulation of ion homoeostasis and volume control in mammalian cells.
2008,
Pubmed
Frank,
The amino-acid sequence of the alkali light chains of rabbit skeletal-muscle myosin.
1974,
Pubmed
Franzetti,
Tetrahedral aminopeptidase: a novel large protease complex from archaea.
2002,
Pubmed
Gagnon,
Characterization of SPAK and OSR1, regulatory kinases of the Na-K-2Cl cotransporter.
2006,
Pubmed
,
Xenbase
Gagnon,
Volume sensitivity of cation-Cl- cotransporters is modulated by the interaction of two kinases: Ste20-related proline-alanine-rich kinase and WNK4.
2006,
Pubmed
,
Xenbase
Gagnon,
Functional insights into the activation mechanism of Ste20-related kinases.
2011,
Pubmed
,
Xenbase
Gagnon,
Molecular physiology of SPAK and OSR1: two Ste20-related protein kinases regulating ion transport.
2012,
Pubmed
Geng,
The Ste20 kinases Ste20-related proline-alanine-rich kinase and oxidative-stress response 1 regulate NKCC1 function in sensory neurons.
2009,
Pubmed
Gondzik,
Coupling of epithelial Na+ and Cl- channels by direct and indirect activation by serine proteases.
2012,
Pubmed
Grimm,
SPAK isoforms and OSR1 regulate sodium-chloride co-transporters in a nephron-specific manner.
2012,
Pubmed
Kay,
The importance of being proline: the interaction of proline-rich motifs in signaling proteins with their cognate domains.
2000,
Pubmed
Kelley,
Protein structure prediction on the Web: a case study using the Phyre server.
2009,
Pubmed
Kleyman,
ENaC at the cutting edge: regulation of epithelial sodium channels by proteases.
2009,
Pubmed
Kr zel,
Coordination of heavy metals by dithiothreitol, a commonly used thiol group protectant.
2001,
Pubmed
Lin,
Impaired phosphorylation of Na(+)-K(+)-2Cl(-) cotransporter by oxidative stress-responsive kinase-1 deficiency manifests hypotension and Bartter-like syndrome.
2011,
Pubmed
Ma,
IDPicker 2.0: Improved protein assembly with high discrimination peptide identification filtering.
2009,
Pubmed
MacCoss,
Shotgun identification of protein modifications from protein complexes and lens tissue.
2002,
Pubmed
Martinez,
Obesity and altered glucose metabolism impact HDL composition in CETP transgenic mice: a role for ovarian hormones.
2012,
Pubmed
McCormick,
The WNKs: atypical protein kinases with pleiotropic actions.
2011,
Pubmed
McCormick,
A SPAK isoform switch modulates renal salt transport and blood pressure.
2011,
Pubmed
Park,
Regulation of NKCC2 activity by inhibitory SPAK isoforms: KS-SPAK is a more potent inhibitor than SPAK2.
2013,
Pubmed
,
Xenbase
Piechotta,
Cation chloride cotransporters interact with the stress-related kinases Ste20-related proline-alanine-rich kinase (SPAK) and oxidative stress response 1 (OSR1).
2002,
Pubmed
Piechotta,
Characterization of the interaction of the stress kinase SPAK with the Na+-K+-2Cl- cotransporter in the nervous system: evidence for a scaffolding role of the kinase.
2003,
Pubmed
,
Xenbase
Ponce-Coria,
Regulation of NKCC2 by a chloride-sensing mechanism involving the WNK3 and SPAK kinases.
2008,
Pubmed
,
Xenbase
Puente,
A genomic analysis of rat proteases and protease inhibitors.
2004,
Pubmed
Rafiqi,
Role of the WNK-activated SPAK kinase in regulating blood pressure.
2010,
Pubmed
Ranaldi,
Intracellular distribution of labile Zn(II) and zinc transporter expression in kidney and MDCK cells.
2002,
Pubmed
Richardson,
Activation of the thiazide-sensitive Na+-Cl- cotransporter by the WNK-regulated kinases SPAK and OSR1.
2008,
Pubmed
Rossier,
Activation of the epithelial sodium channel (ENaC) by serine proteases.
2009,
Pubmed
San-Cristobal,
Angiotensin II signaling increases activity of the renal Na-Cl cotransporter through a WNK4-SPAK-dependent pathway.
2009,
Pubmed
,
Xenbase
Schwacke,
Network modeling reveals steps in angiotensin peptide processing.
2013,
Pubmed
Tamura,
Noncompetitive, reversible inhibition of aminoacylase-1 by a series of L-alpha-hydroxyl and L-alpha-fluoro fatty acids: ligand specificity of aspergillus oryzae and porcine kidney enzymes.
2000,
Pubmed
Terker,
Sympathetic stimulation of thiazide-sensitive sodium chloride cotransport in the generation of salt-sensitive hypertension.
2014,
Pubmed
Timmer,
Structural and kinetic determinants of protease substrates.
2009,
Pubmed
Villa,
Structural insights into the recognition of substrates and activators by the OSR1 kinase.
2007,
Pubmed
Wilk,
Purification, characterization, and cloning of a cytosolic aspartyl aminopeptidase.
1998,
Pubmed
Williamson,
The structure and function of proline-rich regions in proteins.
1994,
Pubmed
Wolf,
Aminopeptidase A: a key enzyme in the intrarenal degradation of angiotensin II.
1997,
Pubmed
Yang,
SPAK-knockout mice manifest Gitelman syndrome and impaired vasoconstriction.
2010,
Pubmed
Yates,
Method to correlate tandem mass spectra of modified peptides to amino acid sequences in the protein database.
1995,
Pubmed
Yokoyama,
Identification of yeast aspartyl aminopeptidase gene by purifying and characterizing its product from yeast cells.
2006,
Pubmed
