Phosphoproteome Mapping of Cardiomyocyte Mitochondria in a Rat Model of Heart Failure

Supplementary Data

Phosphoproteome mapping of cardiomyocyte mitochondria in a rat model of heart failure Molecular and Cellular Biochemistry Francesco Giorgianni, M. Usman Khan, Karl T. Weber, Ivan C. Gerling, Sarka Beranova-Giorgianni Corresponding author: Sarka Beranova-Giorgianni, University of Tennessee Health Science Center, [email protected]

Supplementary Table 1. Phosphopeptides, sites, and proteins characterized in rat heart mitochondria from ALDOST and Control samples. The entries are listed in the same order as in Table 1 in the main body of the article. Data for ALDOST (two biological replicates) and Control are shown here for qualitative purposes; no quantitative comparison was performed.

No UniProt Protein Name Peptide Sequence1 Site Site Location ALDOST ALDOST CONTROL

Accession Probability Pool 1 Pool 2 Pool 1

Number [%]2,3 1 P26284 Pyruvate dehydrogenase E1 YGMGTsVER S232 95.8 + + +

component subunit alpha , YGmGTsVER S232 - + - S293 99.9 + + + somatic form, YHGHsMSDPGVSYR S293 - + - mitochondrial YHGHsmSDPGVSYR S293/S300 99.9/99.4 + - + YHGHsMSDPGVsYR 2 Q00972 [3-methyl-2-oxobutanoate StSATDTHHVELAR T32 33.3 + - +

dehydrogenase [lipoamide]]

kinase, mitochondrial

(BCKD-kinase) 3 P15650 Long-chain specific acyl- IFsSEHDIFR S54 99.4 + + + CoA dehydrogenase ,

mitochondrial (LCAD) 4 Q5M9I5 Cytochrome b c-1 complex sQTEEDCTEELFDFLH S59 83.5 - - +

subunit 6 AR T61 72.2 + - -

VSSRSQtEEDCTEELF

DFLHAR

5 P15999 ATP synthase subunit S521 99.4 + + +

alpha, mitochondrial FESAFLSHVVSQHQs

LLGNIR

6 Q75Q39 Mitochondrial import AsPALGSGPDGSGDS S94 100 + + -

receptor subunit TOM70 LEMSSLDR 7 Q9WVA1 Mitochondrial import inner SKPVFSESLsD S96 99.3 + + +

membrane translocase

subunit Tim8 A 8 P34058 Heat shock protein HSP-90 IEDVGsDEEDDSGKD S255 99.4 - - +

beta KK 9 P11507 Sarcoplasmic/endoplasmic EFDELSPsAQR S663 100 + + +

reticulum calcium ATPase

2 (SERCA2) 10 P61016 Cardiac phospholamban RAsTIEMPQQAR S16 94.5 + + +

RAstIEMPQQAR S16/T17 100/100 + + + 11 P23562 Band 3 anion transport VLEIPDRDsEEELEHV S18 100 + + + protein IEQIAYR 12 P19357 Solute carrier family 2, RTPsLLEQEVKPSTEL S488 22.4 + + +

facilitated glucose EYLGPDEND

transporter member 4

(GLUT-4) 13 P08050 Gap junction alpha-1 QAsEQNWANYSAE S306 100 + + +

protein (connexin-43) QNR S328 25.0 + + + S328/S330 16.7/16.7 + - - MGQAGSTIsNSHAQ S365 79.2 - - + PFDFPDDNQNAK S365/S368/S369 55.9/94.1/ + - + MGQAGSTIsNsHAQ 94.1 PFDFPDDNQNAK

VAAGHELQPLAIVDQ

RPSsRAssR 14 P09605 Creatine kinase S-type, LGYILTCPsNLGTGLR S319 100 + + +

mitochondrial (S-MtCK) 15 Q05962 ADP/ATP translocase 1 LLLQVQHAsK S42 100 + - -

(ANT 1) YFPtQALNFAFK T84 100 + + + Y195 100 + + + AAYFGVyDTAK 16 O08589 Phospholemman TGEPDEEEGTFRSsIR S83 99.8 + - - 17 P02563 Myosin-6 RNtLLQAELEELR T1680 100 + - - 18 P60905 DNAJ homolog subfamily sLSTSGESLYHVLGLD S8 48.4 + + +

C member 5 K 19 P04692 Tropomyosin alpha-1 chain AISEELDHALNDMTsI S283 99.8 + + + 20 Q9Z2L0 Voltage-dependent anion- LTFDSSFsPNTGKK S104 88.0 + + -

selective channel protein 1 DVFtKGYGFGLIK T19 99.7 + + -

(VDAC1) 21 Q9ER34 Aconitate hydratase, WVVIGDENYGEGsS S669 43.0 + - -

mitochondrial REHAALEPR 22 B1WBW4 Armadillo repeat- sAEDLTEGSYDAILSA S43 100 + + -

containing protein 10 EQLEK 23 Q68FT1 Ubiquinone biosynthesis YTDQsGEEEEDYESE S80 91.0 + + +

protein COQ9, EQIQHR

mitochondrial 24 P53987 Monocarboxylate SKEsLQEAGKSDANT S213 81.9 + + -

transporter 1 DLIGGSPK 25 P11960 2-oxoisovalerate IGHHsTSDDSSAYR S333 96.7 - + +

dehydrogenase subunit sVDEVNYWDK S343 100 - - + S343 + - + alpha, mitochondrial sVDEVNYWDKQDH S333/S343 + + + PISR

IGHHsTSDDSSAYRsV

DEVNYWDK 26 B0LPN4 Ryanodine receptor 2 RIsQTSQVSIDAAHG S2798 32.6 - + -

YSPR 1 Phosphorylation site(s) are indicated with lower-case letters. Lower-case m denotes oxidized methionine. 2 Phosphosite localization probabilities were calculated with the phosphoRS module of Proteome Discoverer (Taus et al, (2011) J Proteome Res 10:5354). 3 For phosphosites characterized in multiple samples, the highest site localization probability is shown.