CKDdb-logoCKDdb - Study ID Exp24761003c

PubMed ID24761003
AuthorsStodkilde L, Palmfeldt J, Nilsson L, Carlsen I, Wang Y, Norregaard R, Frokiaer J.
TitleProteomic identification of early changes in the renal cytoskeleton in obstructive uropathy.
JournalAm J Physiol Renal Physiol. 2014 Jun 15;306(12):F1429-41.
AbstractBilateral ureteral obstruction (BUO) is associated with renal damage and impaired ability to concentrate urine and is known to induce alterations in an array of kidney proteins. The aim of this study was to identify acute proteomic alterations induced by BUO. Rats were subjected to BUO for 2, 6, or 24 h. Mass spectrometry-based proteomics was performed on the renal inner medulla, and protein changes in the obstructed group were identified. Significant changes were successfully identified for 109 proteins belonging to different biological classes. Interestingly, proteins belonging to the cytoskeleton and proteins related to cytoskeletal regulation were found to be biologically enriched in BUO using online-accessible tools. Western blots confirmed the selected results, demonstrating acute downregulation of proteins belonging to all three cytoskeletal components. The microfilament protein beta-actin and the intermediate filament proteins pankeratin and vimentin were all downregulated. beta-Tubulin, an important microtubular protein, was found to be significantly downregulated after 24 h. Also, there was significant upregulation of cofilin, an actin-binding protein known to be upregulated in other nephropathy models. Furthermore, both upregulation and downregulation of cytoskeletal motor and regulatory proteins were observed. These findings were confirmed by immunohistochemistry, which clearly showed alterations in labeling in the inner medulla. Interestingly, we were able to confirm selected results in mpkCCD cells exposed to mechanical stretch. Our findings add to the knowledge of BUO-induced acute changes in the renal cytoskeleton and suggest that these molecular changes are partly mediated by increased stretch of the cells during obstruction.

Sample characteristics

SpeciesTissue / SourceCompartmentDiseaseNDetection method
Rattus norvegicuskidneymedullaureteral_obstruction18LC-MS/MS

Sample description and preparation

N (case)9
N (control)9
Disease (case)obstructive uropathy
Disease (control)healthy, sham operated
Disease inductionocclude the ureters (BUO), 24hrs, soluble fraction
Preparation specific methodiTRAQ labelling
Digestin-solution, trypsin

Molecule list

Molecule IDextExternal IDGeneNameRegulation (case/control)Scores
A9639 RS21_RATRps2140S ribosomal protein S21ratio: 3.51
A9580 RLA2_RATRplp2, Rp260S acidic ribosomal protein P2ratio: 2.1
A0451 GRP78_RATHspa5, Grp7878 kDa glucose-regulated proteinratio: 1.52
A0429XACON_RATAco2, Aco2Aconitate hydrataseratio: 0.74
A790B ACBP_RATDbi, Dbi, LOC679040Acyl-CoA-binding proteinratio: 0.42
A0240 CRK_RATCrk, Crko, CrkAdapter molecule crkratio: 1.42
A0802XAKA12_RATAkap12, Akap12, 322A-kinase anchor protein 12ratio: 0.8
A5625XA1AG_RATOrm1Alpha-1-acid glycoproteinratio: 0.4
A295F AMGO3_RATAmigo3, Ali3Amphoterin-induced protein 3ratio: 0.56
A4575XANXA4_RATAnxa4, Anx4, Anxa4Annexin A4ratio: 0.7
A269A CAPR1_RATCaprin1, Gpiap1, Rng105Caprin-1ratio: 1.71
A5965 CAH3_RATCa3Carbonic anhydrase 3ratio: 0.45
A4725 COTL1_RATCotl1, ClpCoactosin-like proteinratio: 1.53
A894BXCOPB_RATCopb1, CopbCoatomer subunit betaratio: 0.87
A0999 COF1_RATCfl1, Cfl1Cofilin-1ratio: 1.11
A6087XCNDP2_RATCndp2, Cndp2Cytosolic non-specific dipeptidaseratio: 0.69
A0442XDPYL2_RATDpysl2Dihydropyrimidinase-related protein 2ratio: 0.73
A0656XDCTN2_RATDctn2Dynactin subunit 2ratio: 0.5
A4778 EMAL2_RATEml2, Emap2, Eml2Echinoderm microtubule-associated protein-like 2ratio: 0.5
A3804 EF2_RATEef2, Eef2, Dapk3Elongation factor 2ratio: 1.08
A6426 ENOPH_RATEnoph1, MasaEnolase-phosphatase E1ratio: 1.46
A972BXFABP4_RATFabp4, Fabp4, Fabp4Fatty acid-binding proteinratio: 1.47
A977B FABPH_RATFabp3Fatty acid-binding proteinratio: 1.11
A1467XFIBA_RATFga, Fga, FgaFibrinogen alpha chainratio: 2.03
A1493 FIBB_RATFgb, FgbFibrinogen beta chainratio: 2.49
A3616 ENOG_RATEno2, Eno-2, Eno2Gamma-enolaseratio: 1.5
A1961XGSTP1_RATGstp1, Gstp1Glutathione S-transferase Pratio: 0.37
A4856XHSPB1_RATHspb1, Hsp27, Hspb1Heat shock protein beta-1ratio: 0.49
A2547 ROA3_RATHnrnpa3, Hnrpa3, Hnrnpa3Heterogeneous nuclear ribonucleoprotein A3ratio: 3.08
A1611XHRG_RATHrg, Hrg1, HrgHistidine-rich glycoproteinratio: 1.47
A3786 K1C13_RATKrt13, Ka13Keratinratio: 2.03
A0808XMOES_RATMsn, MsnMoesinratio: 0.8
A8376 A1I3_RATA1i3, Mug1, Mug1Alpha-1-inhibitor 3ratio: 0.59
A3166 MYH9_RATMyh9, LOC100911597, Myh9Myosin-9ratio: 0.7
A6256XDDAH2_RATDdah2N(G)ratio: 1.16
A0806 NHRF1_RATSlc9a3r1, Nherf, Nherf1Na(+)/H(+) exchange regulatory cofactor NHE-RF1ratio: 0.39
A2536XNUCL_RATNcl, Nuc, NclNucleolinratio: 1.71
A5418 NP1L4_RATNap1l4Nucleosome assembly protein 1-like 4ratio: 2.27
A3883 PALM_RATPalm, PalmParalemmin-1ratio: 0.39
A7375 PGM1_RATPgm1, Pgm1, Pgm1Phosphoglucomutase-1ratio: 1.8
A797D PHLB1_RATPhldb1, Ll5, Phldb1Pleckstrin homology-like domain family B member 1ratio: 0.7
A277CXPDC6I_RATPdcd6ip, Aip1, Pdcd6ipProgrammed cell death 6-interacting proteinratio: 0.52
A838A PDCD4_RATPdcd4, Dug, Pdcd4Programmed cell death protein 4ratio: 0.25
A8971XPSME1_RATPsme1, Psme1Proteasome activator complex subunit 1ratio: 0.35
A8383XAMBP_RATAmbp, ItilProtein AMBPratio: 1.47
A0323 RAP1A_RATRap1a, Krev1, Rap1aRas-related protein Rap-1Aratio: 1.62
A463C SBP1_RATSelenbp1, Sbp, Selenbp1Selenium-binding protein 1ratio: 0.3
A8492 SPA3K_RATSerpina3k, Spin2bSerine protease inhibitor A3Kratio: 0.59
A0353 SPA3L_RATSerpina3l, Spin2aSerine protease inhibitor A3Lratio: 0.57
A8642 2ABA_RATPpp2r2aSerine/threonine-protein phosphatase 2A 55 kDa regulatory subunit B alpha isoformratio: 0.71
A643CXTRFE_RATTf, Tf, SrprbSerotransferrinratio: 0.98
A1581XALBU_RATAlbSerum albumin ( Precursor)ratio: 0.72
A5153XSPTN2_RATSptbn2, Spnb3, Sptbn2Spectrin beta chainratio: 0.64
A2010XGRP75_RATHspa9, Grp75, Hspa9aStress-70 proteinratio: 0.62
A9106 THIO_RATTxn, Txn1Thioredoxinratio: 0.67
A652C TXNL1_RATTxnl1, Trp, Txnl1Thioredoxin-like protein 1ratio: 0.5
A9969 KNT1_RATMap1T-kininogen 1ratio: 3.92
A9970 KNT2_RAT T-kininogen 2ratio: 4.62
A648CXTTHY_RATTtr, TtTransthyretinratio: 0.9
A5195XTBB5_RATTubb5, RT1-N2, 2310ex4-5Tubulin beta-5 chainratio: 0.62
A400EXD3ZW65Tpd52l2Tumor protein D52-like 2ratio: 1.65
A472E UP1_RAT Urinary protein 1ratio: 1.83
A0850XVIME_RATVim, VimVimentinratio: 1.04
A7357XPEPD_RATPepd, Pepd, PepdXaa-Pro dipeptidaseratio: 0.62

Compile date 08-10-2018© iMODE-CKD consortium