Rubisco peptides were analyzed by MS on an LTQ Orbitrap XL system, and peak areas of the precursor ions (mass tolerance ≤ 10 ppm) were manually quantified from two biological replicates treated and untreated with hSIRT3 using the peak detection function of the Xcalibur program (Thermo Fisher Scientific). Retention times of the precursor ions are indicated in parentheses after the respective peak area. Ratio = peak area (+hSIRT3)/peak area (−hSIRT3); n = 2. n.d., Not detected.
Lys (K) Side Chain Site | Peptide | Precursor Mass | z | Peak Area Replicate (1) | Peak Area Replicate (2) | Ratio (1) | Ratio (2) | ||
−hSIRT3 | +hSIRT3 | −hSIRT3 | +hSIRT3 | ||||||
K14 | K.ASVGFK(ac)AGVK.E | 503.2893 | 2 | 2,024,658 (38.68) | 1,743,313 (39.43) | 4,323,830 (44.89) | 1,994,744 (45.89) | 0.86 | 0.46 |
K18 | K.AGVK(ac)EYK.L | 418.7293 | 2 | 6,042 (18.87) | n.d. | 181,431 (21.57) | 78,107 (22.07) | 0.00 | 0.43 |
K21 | K.EYK(ac)LTYYTPEYETK.D | 935.4443 | 2 | 540,204 (55.79) | n.d. | 166,379 (61.00) | 92,189 (61.34) | 0.00 | 0.55 |
K146 | R.IPPAYTK(ac)TFQGPPHGIQVER.D | 760.0713 | 3 | 105,885 (48.06) | 97,448 (47.75) | n.d. | n.d. | 0.92 | n.d. |
K175 | K.YGRPLLGC(ca)TIKPK(ac)LGLSAK.N | 705.4099 | 3 | 371,434 (44.7) | n.d. | n.d. | n.d. | 0.00 | n.d. |
K252 | K.GHYLN(da)ATAGTC(ca)EEMIK(ac)R.A | 997.459 | 2 | 2,139,141 (22.98) | 1,454,392 (23.57) | 271,229 (30.3) | 48,605 (30.52) | 0.68 | 0.18 |
K252 | K.GHYLNATAGTC(ca)EEMIK(ac)R.A | 664.9806 | 3 | 147,742 (56.13) | n.d. | n.d. | n.d. | 0.00 | n.d. |
K252 | K.SQAETGEIKGHYLN(da)A(TAGTCca)EEMIK(ac)R.A | 735.0985 | 4 | 6,310,874 (31.29) | 3,437,305 (32.02) | 1,862,689 (36.36) | 1,457,631 (36.41) | 0.54 | 0.78 |
K252 | K.SQAETGEIKGHYLNATAGTC(ca)EEMIK(ac)R.A | 979.4679 | 3 | 114,592 (55.33) | n.d. | 71,688 (51.17) | 29,682 (51.02) | 0.00 | 0.41 |
K252 | K.SQAETGEIKGHYLN(da)ATAGTC(ca)EEMIK(ac)R.A | 979.7959 | 3 | 52,038 (40.01) | 14,805 (40.43) | 79,698 (44.37) | 25,745 (43.79) | 0.28 | 0.32 |
K316 | R.VLAK(ac)ALR.L | 406.7712 | 2 | 560,705 (22.32) | 447,572 (22.91) | 7,311,999 (25.42) | n.d | 0.80 | 0.00 |
K356 | R.DDYVEK(ac)DR.S | 541.2435 | 2 | 931,502 (21.57) | 8,349 (21.32) | n.d. | n.d. | 0.01 | n.d. |
K356 | R.ESTLGFVDLLRDDYVEK(ac)DR.S | 771.3849 | 3 | 10,723,480 (51.75) | 3,560,598 (52.44) | 1,457,839 (52.42) | 244,713 (53.04) | 0.33 | 0.17 |
K356 | R.ESTLGFVDLLRDDYVEK(ac)DR.S | 1,156.5739 | 2 | 3,375,184 (87.55) | n.d. | 1,095,145 (91.52) | 286,040 (90.31) | 0.00 | 0.26 |