Figure s1 general experimental procedure
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Figure S1. General experimental procedure. WT. Nuclei prep/Acid Extraction. H2B. Intentisity. Step 1. Propionylation Step 2. Trypsin digestion Step 3. Propionylation. Intentisity. ΔEZL2. H4. H2A. H3. ΔTXR1. RT. M/Z. Equal mixture of L&H histones. N15 WT.

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Figure S1. General experimental procedure

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Figure s1 general experimental procedure

Figure S1. General experimental procedure

WT

Nuclei prep/Acid Extraction

H2B

Intentisity

Step 1. Propionylation

Step 2.

Trypsin digestion

Step 3.

Propionylation

Intentisity

ΔEZL2

H4

H2A

H3

ΔTXR1

RT

M/Z

Equal mixture

of L&H histones

N15 WT

RP-18 HPLC separation

NanoLC-MS/MS analysis


A h2a variants

Figure S2. Histone variants, sequence coverage, and their PTMs

a. H2A variants

H2A   --------STTGKGGKAKG-KTASSKQVSRSARAGLQFPVGRISRFLKNGRYS-ERIGTG 50

H2A.X  --------STTGKGGKAKG-KTASSKQVSRSARAGLQFPVGRISRFLKHGRYS-ERVGTG 50

H2A.Z  AGGKGGKGGKGGKGGKVGGAKNKKTPQ-SRSYKAGLQFPVGRIHRFLKGRVSAKNRVGAT 59

H2A APVYLAAVLEYLAAEVLELAGNAAKDNKKTRIVPRHILLAIRNDEELNKLMANTTIADGG 110

H2A.X APVYLAAVLEYLAAEVLELAGNAAKDNKKTRIVPRHILLAIRNDEELNKLMANTTIADGG 110

H2A.Z AAVYAAAILEYLTAEVLELAGNASKDFKVRRITPRHLLLAIRGDEEL-DILIKATIAGGG 118

H2A   VLPNINPMLLPSKTKKS-----TEPEH  132

H2A.X  VLPNINPMLLPSKSKKTESRGQASQDL  137

H2A.Z  VIPHIHKALLGKHSTKNRSSAKTAEPR  145

Sequence coverage: 60%

# of PTMs identified (17 PTMs):

H2A.X/H2A.1:K5Ac, K8Ac, K10Ac, K12Ac, S1/T2/T3Phos*, R33Me1, R36Me2, K39Form, K42Me1, R92Me1

H2A.Z:K4Ac,K7Ac,K10Ac,K13Ac,K16Ac,K21Ac,K140Ac

# of PTMs quantified (13 PTMs):

H2A.X/H2A.1:K5Ac, K8Ac, K10Ac, K12Ac, K39Form, R92Me1

H2A.Z:K4Ac,K7Ac,K10Ac,K13Ac,K16Ac,K21Ac,K140Ac

*Localization is difficult due to incomplete fragment ion information


B h2b variants

b. H2B variants

H2B1 APKKAPAAAAEKKVKKAPTTEKKNKKKRSETFAIYIFKVLKQVHPDVGISKKAMNIMNSF 60

H2B2 APKKAPAATTEKKVKKAPTTEKKNKKKRSETFAIYIFKVLKQVHPDVGISKKAMNIMNSF 60

H2B1 INDSFERIALESSKLVRFNKRRTLSSREVQTAVKLLLPGELARHAISEGTKAVTKFSSST 120

H2B2 INDSFERIALESSKLVRFNKRRTLSSREVQTAVKLLLPGELARHAISEGTKAVTKFSSSS 120

H2B1 N 121

H2B2 N 121

Sequence coverage: 68%

# of PTMs identified (12 PTMs):

H2B-1: A1Me1, K3Me1/Me2/Me3, K4Ac, K115Ac, (S117-T120)Phos*

H2B-2: A1Me1, K3Me1/Me2/Me3, K4Ac

# of PTMs quantified (10 PTMs):

H2B-1: A1Me1, K3Me1/Me2/Me3, K4Ac, K115Ac, (S117-T120)Phos*

H2B-2: K3Me2/Me3, K4Ac

*Localization is difficult due to incomplete fragment ion information


Figure s1 general experimental procedure

c. H3 variants

H3

H3.3

H3.4

H3

H3.3

H3.4

H3

H3.3

H3.4

ARTKQTARKSTGAKAPRKQLASKAARKSAPATGGIKKPHRFRPGTVALREIRKYQKSTDL 60

ARTKQTARKSTGVKAPRKQLATKAARKSAPVSGGVKKPHKFRPGTVALREIRKYQKTTDL 60

ARTKQTARKSTSIKAPRKQLAAKAARKSAPISGGIKKPHKFRPGTVALREIRKYQKTTDL 60

LIRKLPFQRLVRDIAHEFKAELRFQSSAVLALQEAAEAYLVGLFEDTNLCAIHARRVTIM 120

LIRKLPFQRLVRDIAMEMKSDIRFQSQAILALQEAAEAYLVGLFEDTNLCAIHARRVTIM 120

LIRKLPFQRLVRDIAMEMKSDIRFQSQAILALQEAAEAYLVGLFEDTNLCAIHARRVTIM 120

TKDMQLARRIRGERF 135

TKDLHLARRIRGERF 135

TKDLHLARRIRGERF 135

Sequence coverage: 76%

# of PTMs identified (32 PTMs):

H3: K4Me1/Me2/Me3, K9Ac, K14Ac, K14Me1, K18Ac, K23Ac, K23Me1/Me2/Me3, K23Form, K27Me1/Me2/Me3, K27Ac, K36Me1/Me2, R40Me1, R49Me1, K56Me1, K79Me1, R83Me1, R128Me1

H3.3: K9Ac, K14Ac, K14Form, K18Ac, K23Ac, K23Form, K56Ac, K79Me1

# of PTMs quantified (25 PTMs):

H3: K4Me1/Me3, K9Ac, K14Ac, K18Ac, K23Ac, K23Me1/Me2/Me3, K23Form, K27Me1/Me2/Me3, K27Ac, K36Me1, K56Me1, R83Me1, R128Me1

H3.3: K9Ac, K14Ac, K14Form, K18Ac, K23Ac, K23Form, K56Ac


Figure s1 general experimental procedure

d. H4

AGGKGGKGMGKVGAKRHSRKSNKASIEGITKPAIRRLARRGGVKRISSFIYDDSRQVLKS 60

FLENVVRDAVTYTEHARRKTVTAMDVVYALKRQGRTLYGFGG 102

Sequence coverage: 97%

# of PTMs identified (11 PTMs):

H4: K4Ac, K7Ac, K7Ub, K11Ac, K15Ac, R35Me1, R55Me1, R67Me1, R77Me1, K91Me1, R92Me1

# of PTMs quantified (7 PTMs):

H4: K4Ac, K7Ac, K11Ac, K15Ac, R55Me1, R77Me1, K91Me1


Figure s1 general experimental procedure

Figure S3. All PTMs identified by tandem MS

H2A.X-K5Ac

1STTGKGGKAK10

[M-2H2O+2H]2+

b8

b9

y2

y1

y3

b6

y4

b7

y5

M/Z: 572.8141, Charge: 2+, Mass Error: 0ppm, Ion Score: 23


Figure s1 general experimental procedure

H2A.X-K5AcK8AcK10AcK12Ac

1STTGKGGKAKGKTASSKQVSR21

[M-2H2O+2H]2+

y7

b14-H2O

y11

y9

y8

y6

b17-H2O

y4

y5

y3

b13

b15-H2O

b18

b16

b16-H2O

y13

y12

M/Z: 1172.628, Charge: 2+, Mass Error: 1ppm, Ion Score: 54


Figure s1 general experimental procedure

H2A.X-K5AcK8AcK10Ac

1STTGKGGKAKGKTASSKQVSR21

[M-2H2O+2H]2+

y19-NH3

y7

b20

y4

y6

b7-NH3

y8-H2O

y2

y5

y3

y9

y8

y10

b8

b12

M/Z: 786.7592, Charge: 3+, Mass Error: 0ppm, Ion Score: 34


Figure s1 general experimental procedure

H2A.X-K5AcK8AcK12Ac

1STTGKGGKAKGKTASSKQVSR21

[M-2H2O+2H]2+

y11

y7

b12

b14-H2O

y9

y6

y8

b17-H2O

b18-H2O

y12

b15

y4

y13

b16-H2O

y3

y5

b18

M/Z: 1179.6361, Charge: 2+, Mass Error: 1ppm, Ion Score: 69


Figure s1 general experimental procedure

H2A.X-K5AcK8Ac

1STTGKGGKAKGKTASSKQVSR21

[M-2H2O+2H]2+

y9

b12

y7

y11

b14-H2O

b10

b15-H2O

y6

y8

y4

b13

y5

y16

y12

y13

b15

b14

M/Z: 1186.6435, Charge: 2+, Mass Error: 1ppm, Ion Score: 101


Figure s1 general experimental procedure

H2A.X-STT(1-3)Phos

1STTGKGGKAKGKTASSKQVSR21

[M-H2O+2H]2+

b10-NH3

b17

b6

y9

y7

b9

y11

b12

b13

b14

b16

b5

y6

b15

M/Z: 1212.6277, Charge: 2+, Mass Error: 1ppm, Ion Score: 43


Figure s1 general experimental procedure

H2A.X-K10AcK12Ac

6GGKAKGKTASSKQVSR21

y11

y9

y7

y8

b14

y6

b7

y4

b5

b14-H2O

b12-H2O

y3

y5

y10

b13

b15

b12

y2

b10

b11

b15-H2O

b6

M/Z: 893.4959, Charge: 2+, Mass Error: 2ppm, Ion Score: 72


Figure s1 general experimental procedure

H2A.X-K10Ac

6GGKAKGKTASSKQVSR21

[M-2H2O+2H]2+

y7

y9

b14

y4

y11

y6

y5

b15

b7

y10

b13

b14-H2O

b6

y2

y3

b10

b12

y12

M/Z: 900.504, Charge: 2+, Mass Error: 2ppm, Ion Score: 79


Figure s1 general experimental procedure

H2A.X-K12Ac

6GGKAKGKTASSKQVSR21

[M-H2O+2H]2+

y7

y9

y4

b6-NH3

b14

y11

b15

y5

b10

b12

b6

b14-NH3

b5

y6

b12-NH3

b11

b13

y10

y13

M/Z: 912.4688, Charge: 2+, Mass Error: 2ppm, Ion Score: 58


Figure s1 general experimental procedure

H2A.X-R33Me1

25AGLQFPVGR33

[M-H2O+2H]2+

y5

y6

y4

b5

b3

y2

b4

b7

y1

b6

y3

y8

M/Z: 514.2709, Charge: 2+, Mass Error: 0ppm, Ion Score: 30


Figure s1 general experimental procedure

H2A.X-R36Me2

34ISRFLKHGR42

[M-H2O+2H]2+

y8,2+

y3

y4

y6

y5

y8

y7,2+

b7

b3-H2O

y2

b6

b5

y7

b8

M/Z: 627.3735, Charge: 2+, Mass Error: -6ppm, Ion Score: 43


Figure s1 general experimental procedure

H2A.X-R42Me1

37FLKHGR42

y3

y5,2+

y4

y5

y2

[M-H2O+2H]2+

b1

b4

b3

b2

b5

M/Z: 448.2434, Charge: 2+, Mass Error: 0ppm, Ion Score: 39


Figure s1 general experimental procedure

H2A.X-K39Form

37FLKHGR42

y5,2+

y4

[M-2H2O+2H]2+

y5

y3

y2

y1

b5-NH3

b4,2+

b4-NH3

b1

b2

b5

M/Z: 421.2374, Charge: 2+, Mass Error: 0ppm, Ion Score: 36


Figure s1 general experimental procedure

H2A.X-R92Me1

86HILLAIR92

b5

b4

b3

b2

[M-H2O+2H]2+

b6

y4

y5

y3

y6

M/Z: 917.5589, Charge: 1+, Mass Error: 2ppm, Ion Score: 53


Figure s1 general experimental procedure

H2A.Z-K7AcK10Ac

1AGGKGGKGGKGGK13

y9

y11,2+

y10,2+

y8

b7

b10

b3

y3

b11

y11

b5

y7

b4

y5

b9

b12

y4

M/Z: 571.8189, Charge: 2+, Mass Error: 3ppm, Ion Score: 58


Figure s1 general experimental procedure

H2A.Z-K4AcK7AcK10Ac

1AGGKGGKGGKGGK13

y9

y11,2+

y8

y6

y10,2+

b10

y3

b7

y7

b4

b5

b11

b12

y4

b6

y5

y11

b9

b8

M/Z: 592.8227, Charge: 2+, Mass Error: 0ppm, Ion Score: 68


Figure s1 general experimental procedure

H2A.Z-K4AcK7AcK10AcK13Ac

1AGGKGGKGGKGGKGGK16

y9

y12

y6

y14,2+

b13

b10

y11

b7

y8

y10

y5

y3

b14

b4

b15

y4

b11

b12

y14

b5

b9

M/Z: 734.897, Charge: 2+, Mass Error: 0ppm, Ion Score: 77


Figure s1 general experimental procedure

H2A.Z-K4AcK7AcK10AcK13AcK16Ac

1AGGKGGKGGKGGKGGKVGGAK21

y14

y14,2+

y11

b17

y8

y19,2+

b16

y17

y4

b13

y5

b7

y17,2+

y13

b18

y10

y9

y15

y12

y7

b11

b20

b12

b5

b4

b19

b15

M/Z: 962.0239, Charge: 2+, Mass Error: 0ppm, Ion Score: 97


Figure s1 general experimental procedure

H2A.Z-K4AcK7AcK10AcK13AcK16AcK21Ac

1AGGKGGKGGKGGKGGKVGGAKNK23

b17,2+

y6

b16,2+

y7

y16,2+

b13

b10

y10

b18,2+

b12

y5

y18,2+

y19,2+

b4

b7

b14

b16

b17

b6

y2

y3

b15

y11

y13

b18

M/Z: 736.4026, Charge: 3+, Mass Error: 2ppm, Ion Score: 91


Figure s1 general experimental procedure

H2A.Z-K7AcK10AcK13Ac

5GGKGGKGGKGGK16

y14,2+

y9

y6

y10,2+

b3

b9

b6

y8

y3

y7

y5

b4

b11

b5

b10

b7

y4

y10

M/Z: 557.3039, Charge: 2+, Mass Error: 0ppm, Ion Score: 67


Figure s1 general experimental procedure

H2A.Z-K7AcK10AcK13AcK16Ac

5GGKGGKGGKGGKVGGAK21

y11

y8

b13

y14

y4

b6

y15,2+

b9

b16

y14,2+

y10

y5

b3

y13

y7

y9

b12

y12

b14

b5

b15

y3

M/Z: 784.4312, Charge: 2+, Mass Error: 0ppm, Ion Score: 78


Figure s1 general experimental procedure

H2A.Z-K7AcK10AcK13AcK16AcK21Ac

5GGKGGKGGKGGKVGGAKNK23

b6

y7

y13,2+

b9

y16,2+

y5

b12

b14,2+

y2

y3

b10

b17,2+

b3

b13

y10,2+

y7,2+

y14,2+

y10

y8

b5

y12

M/Z: 618.005, Charge: 3+, Mass Error: -2ppm, Ion Score: 61


Figure s1 general experimental procedure

H2A.Z-K13AcK16Ac

8GGKGGKGGKVGGAK21

y11

y8

[M-H2O+2H]2+

b6

y12,2+

b10

y10

y4

b13

b9

y5

y7

y9

b12

b4

b5

b11

y3

M/Z: 621.3515, Charge: 2+, Mass Error: 0ppm, Ion Score: 55


Figure s1 general experimental procedure

H2A.Z-K13AcK16AcK21Ac

8GGKGGKGGKVGGAKNK23

y6

[M-H2O+2H]2+

b10

y10

y14,2+

y13

b6

b14

b13

b9

y9

y8

y3

y13,2+

b15

b11

y11,2+

y12

y2

y11

b12

b4

b5

y7

M/Z: 774.3933, Charge: 2+, Mass Error: 0ppm, Ion Score: 102


Figure s1 general experimental procedure

H2A.Z-K10AcK13AcK16AcK21Ac

8GGKGGKGGKVGGAKNK23

y10

y7

y13

y9

b6

b9

[M-H2O+2H]2+

y3

b13

b15

y13,2+

y8

b3

y12

y12,2+

y6

y11

b10

y14,2+

b11

b4

b14

b5

y4

y5

M/Z: 784.4318, Charge: 2+, Mass Error: 1ppm, Ion Score: 81


Figure s1 general experimental procedure

H2A.Z-K16AcK21Ac

11GGKGGKVGGAKNK23

y10

y7

[M-H2O+2H]2+

b6

y11,2+

y9

b12

y3

b10

y8

b11

y2

b5

y5

b3

y4

b9

y11

M/Z: 621.351, Charge: 2+, Mass Error: -1ppm, Ion Score: 63


Figure s1 general experimental procedure

H2A.Z-K16Ac

14GGKVGGAK21

b7-H2O

y4

y5

b3

b7

y6

b4

y3

y2

M/Z: 725.3801, Charge: 1+, Mass Error: 0ppm, Ion Score: 31


Figure s1 general experimental procedure

H2A.Z-K140Ac

137SSAKTAEPR145

[M-H2O+2H]2+

y2

y5

b7

y3

b7-H2O

y7,2+

y6

y4

b4

b8

y7

y1

b3

M/Z: 494.7563, Charge: 2+, Mass Error: 0ppm, Ion Score: 66


Figure s1 general experimental procedure

H2B-1-A1Me1K3Me1

1APKKAPAAAAEK12

b11

b9

b10

b8

y4

[M-2H2O+2H]2+

b7

[M-H2O+2H]2+

y2

y3

b11,2+

y2-NH3

y1

b6,2+

y5

y8

y7

y9

M/Z: 682.373, Charge: 2+, Mass Error: 0ppm, Ion Score: 52


Figure s1 general experimental procedure

H2B-1-K3Me2

1APKKAPAAAAEK12

b8,2+

b9

y3

b9,2+

b5

b7

y8

b10

b11

y2-H2O

b8

b3-NH3

y9

y11

b2

M/Z: 646.8825, Charge: 2+, Mass Error: 0ppm, Ion Score: 34


Figure s1 general experimental procedure

H2B-1-K3Me3

1APKKAPAAAAEK12

b7,2+

b8,2+

b9

b10

b8

b11

b7

b9,2+

b4

[M-H2O+2H]2+

y2-H2O

y3

y5

y7

b6,2+

y2

y8

M/Z: 653.8902, Charge: 2+, Mass Error: 0ppm, Ion Score: 56


Figure s1 general experimental procedure

H2B-1-A1Me1

1APKKAPAAAAEKK13

y12,2+

y8

y9

y11,2+

b4

b5

y10

b11

y2

b9,2+

[M-H2O+2H]2+

b9

y5

b7

y11

y4

b10

y12

b3

y3

M/Z: 731.9352, Charge: 2+, Mass Error: 0ppm, Ion Score: 47


Figure s1 general experimental procedure

H2B-1-K3Me3K4Ac

1APKKAPAAAAEKK13

b11

b9

b10

b8

b8,2+

y2

y12-H2O

[M-2H2O+2H]2+

b7

y6

y3-H2O

y4

b7,2+

y5

y8

b6,2+

y9

y10

M/Z: 738.9436, Charge: 2+, Mass Error: 1ppm, Ion Score: 53


Figure s1 general experimental procedure

H2B-1-K115Ac

104HAISEGTKAVTKFSSSTN121

b10

[M-2H2O+2H]2+

y14-NH3

y10

y15-NH3

b11

b8

y15

y8

y13

y16,2+

y11

y17

y7

y14

b7

y12

b6

M/Z: 1010.0059, Charge: 2+, Mass Error: 1ppm, Ion Score: 40


Figure s1 general experimental procedure

H2B-1-SSST(117-120)Phos

104HAISEGTKAVTKFSSSTN121

[M-H2O+2H]2+

b10

y13

y15

b3

y8

y14

y17,+2

b11

y7,+2

y10

y11

y13,+2

b3-H2O

y16

y17

M/Z: 1028.9841, Charge: 2+, Mass Error: 1ppm, Ion Score: 38


Figure s1 general experimental procedure

H2B-2-K3Me3

1APKKAPAATTEK12

[M-2H2O+2H]2+

b11

b9

b5

b10

b8

y5

b7

y4

y7

y2

y3

y1

M/Z: 719.3925, Charge: 2+, Mass Error: 1ppm, Ion Score: 44


Figure s1 general experimental procedure

H2B-2-A1Me1

1APKKAPAATTEKK13

b6

y8

y9

b5

b4

y10

y2

b11

y7

y5

b3

b7

b8

b9

b12

b10

M/Z: 761.9465, Charge: 2+, Mass Error: 1ppm, Ion Score: 54


Figure s1 general experimental procedure

H2B-2-K3Me2

1APKKAPAATTEKK13

b11

[M-2H2O+2H]2+

[M-H2O+2H]2+

b8

y2

b9

b5

b12

b10

y7

y4

y8

b7,2+

y3

y12

y5

y9

b2

M/Z: 768.9542, Charge: 2+, Mass Error: 1ppm, Ion Score: 49


Figure s1 general experimental procedure

H2B-2-K3Me3

1APKKAPAATTEKK13

b11

b6

b8

y2

b9

b12

b5

b10

b7

y8

b3

y4

y2-NH3

y9

y3-NH3

b11-NH3

b8,2+

M/Z: 784.4372, Charge: 2+, Mass Error: 1ppm, Ion Score: 58


Figure s1 general experimental procedure

H2B-2-K3Me3K4Ac

1APKKAPAATTEKK13

b11

[M-H2O+2H]2+

[M-2H2O+2H]2+

y2

b8

b10

b9

b12

b5

y7,2+

y2-NH3

y4

y7

y8

y5

b8,2+

y6,2+

y9

y3-NH3

y10

b11-NH3

y3

M/Z: 777.4291, Charge: 2+, Mass Error: 1ppm, Ion Score: 51


Figure s1 general experimental procedure

H3-K4Me1

3TKQTAR8

[M-H2O+2H]2+

y4

[M-2H2O+2H]2+

y5,2+

y1

b2

y3

b3

y5-NH3

y4-H2O

y5

b1

y2

M/Z: 387.7271, Charge: 2+, Mass Error: 0ppm, Ion Score: 33


Figure s1 general experimental procedure

H3-K4Me2

3TKQTAR8

[M-H2O+2H]2+

[M-2H2O+2H]2+

b5

y1

y5,2+

y4

b4

b5-H2O

b2

y5

y3

b3

y2

M/Z: 400.2186, Charge: 2+, Mass Error: 0ppm, Ion Score: 23


Figure s1 general experimental procedure

H3-K4Me3

3TKQTAR8

y5-NH3

[M-H2O+2H]2+

y1

[M-2H2O+2H]2+

b4

b1

b3

y5

b2

y2

y4

y3

M/Z: 401.743, Charge: 2+, Mass Error: 1ppm, Ion Score: 19


Figure s1 general experimental procedure

H3-K9Ac

9KSTGAKAPR17

[M-H2O+2H]2+

[M-2H2O+2H]2+

y8

y2

y3

b1

y7

y6

b4

y4

b7

b2

b5

y5

y1

b3

b6

b8

M/Z: 535.3029, Charge: 2+, Mass Error: -1ppm, Ion Score: 61


Figure s1 general experimental procedure

H3-K14Ac

9KSTGAKAPR17

[M-2H2O+2H]2+

[M-H2O+2H]2+

y2

y7

y3

y7-H2O

y6

y8

y1

y5

b3

b6

b8

y4

M/Z: 507.2902, Charge: 2+, Mass Error: -1ppm, Ion Score: 35


Figure s1 general experimental procedure

H3-K9AcK14Ac

9KSTGAKAPR17

[M-H2O+2H]2+

y8

[M-2H2O+2H]2+

b7

b4,2+

y2

y3

b6

y6

y7

b2

b3

b5

y5

y1

b8

M/Z: 528.2957, Charge: 2+, Mass Error: 0ppm, Ion Score: 60


Figure s1 general experimental procedure

H3-K9AcK14Me1

9KSTGAKAPR17

[M-H2O+2H]2+

[M-2H2O+2H]2+

b7

y8

b6

y2

y3

b1

y8,2+

y6

y5

b8

b2

y7

y1

M/Z: 549.2891, Charge: 2+, Mass Error: -3ppm, Ion Score: 31


Figure s1 general experimental procedure

H3-K18Ac

18KQLASKAAR26

b7

y8

b8

y2

y3

b6-H2O

y5

b4

y8,2+

b1

y4

y1

y7

y6

b6

b5

y7-H2O

b2

M/Z: 563.8324, Charge: 2+, Mass Error: 0ppm, Ion Score: 57


Figure s1 general experimental procedure

H3-K23Ac

18KQLASKAAR26

b7

y8

b8

y3

y2

b6-H2O

b1

y5

y8,2+

b4

y7

y6

y1

b3

b6

y4

b5

b2

y7-H2O

M/Z: 563.8318, Charge: 2+, Mass Error: -1ppm, Ion Score: 54


Figure s1 general experimental procedure

H3-K18AcK23Ac

18KQLASKAAR26

y8

b7

b8

b4

[M-H2O+2H]2+

y5

y2

y3

b1

b6-H2O

y8,2+

y6

b3

y1

y7

y4

b5

b6

b2

M/Z: 556.8272, Charge: 2+, Mass Error: 4ppm, Ion Score: 51


Figure s1 general experimental procedure

H3-K23Me1

18KQLASKAAR26

y8

[M-H2O+2H]2+

b7

b1

b8

y5

y3

b4

b6-NH3

y8,2+

y2

y6

y7

y8-NH3

y1

b2

b6

y7-H2O

y4

M/Z: 585.3263, Charge: 2+, Mass Error: 1ppm, Ion Score: 55


Figure s1 general experimental procedure

H3-K23Me2

18KQLASKAAR26

y8

[M-H2O+2H]2+

b1

y7

y6

b8

y8,2+

y8-NH3

b3

b6

y3

y1

y7,2+

y7-NH3

y5

b7

b2

y2-NH3

y4

y2

M/Z: 556.842, Charge: 2+, Mass Error: -2ppm, Ion Score: 35


Figure s1 general experimental procedure

H3-K23Me3

18KQLASKAAR26

[M-H2O+2H]2+

b8

b5

b1

y8,2+

y6-NH3

y7,2+

y8-NH3

y6,2+

y1

b3

y7

b6

y3

b2

y6

M/Z: 563.8512, Charge: 2+, Mass Error: 1ppm, Ion Score: 22


Figure s1 general experimental procedure

H3-K23Form

18KQLASKAAR26

b7

b8

y2

y3

[M-H2O+2H]2+

b6-H2O

y1

y8,2+

y7

y6

y8

b1

M/Z: 556.8272, Charge: 2+, Mass Error: 4ppm, Ion Score: 25


Figure s1 general experimental procedure

H3-K27AcK36Me2

27KSAPATGGIKKPHR40

y11,2+

y13,2+

y11

b11

y3

y13

y12,2+

y9

b10-NH3

b13

y8

y4

b10

y12

y10

b2

y8,2+

b2-H2O

y9,2+

b3-NH3

y7

y6

y2

b13-H2O

M/Z: 815.4768, Charge: 2+, Mass Error: 2ppm, Ion Score: 75


Figure s1 general experimental procedure

H3-K27Ac

27KSAPATGGIKKPHR40

y11,2+

y11

y3

y13,2+

y8

[M-H2O+2H]2+

y13

y5

y4

y9

b11

y12,2+

y7

b13

b3-H2O

y10

b2-H2O

y12

b9

b10

y6

y9,2+

b2

M/Z: 829.4732, Charge: 2+, Mass Error: 0ppm, Ion Score: 71


Figure s1 general experimental procedure

H3-K27Me1

27KSAPATGGIKKPHR40

y11

y11,2+

y3

y13

y13,2+

[M-H2O+2H]2+

y8

y4

y12,2+

y5

y9

y7

b3-H2O

b13

b1

y12

y9,2+

b2-H2O

y10

b10

b4

y6

b11

b3

b9

M/Z: 843.489, Charge: 2+, Mass Error: 1ppm, Ion Score: 90


Figure s1 general experimental procedure

H3-K27Me2

27KSAPATGGIKKPHR40

[M-H2O+2H]2+

y11

b11

[M-2H2O+2H]2+

y3

y8

y13

b3

b13

y4

y9

y12

b4-NH3

b10

b5

y7

y10

y6

M/Z: 822.4842, Charge: 2+, Mass Error: 1ppm, Ion Score: 88


Figure s1 general experimental procedure

H3-K27Me3

27KSAPATGGIKKPHR40

y11

y3

[M-2H2O+2H]2+

[M-H2O+2H]2+

y8

y4

y13

y9

b13

b11

y7

b10

y12

y5

b6

b7

b9

b8

y6

y10

b3

M/Z: 829.4902, Charge: 2+, Mass Error: -1ppm, Ion Score: 38


Figure s1 general experimental procedure

H3-K27Me1K36Me1

27KSAPATGGIKKPHR40

y11

y11,2+

y3

y13,2+

y13

y4

y5

y8

[M-H2O+2H]2+

y9

y12,2+

y7

[M-2H2O+2H]2+

b13

b1

y12

b2

y8,2+

y6

y10

b10

b11-H2O

y2

M/Z: 850.4978, Charge: 2+, Mass Error: 2ppm, Ion Score: 88


Figure s1 general experimental procedure

H3-K27Me1R40Me1

27KSAPATGGIKKPHR40

y11,2+

y11

y13

y3

y13,2+

[M-H2O+2H]2+

y8

[M-2H2O+2H]2+

y4

y12,2+

y5

y7

b1

y9

b3-H2O

y9,2+

b9

b13

y12

b2-H2O

b3

y2

y10

M/Z: 850.4975, Charge: 2+, Mass Error: 1ppm, Ion Score: 99


Figure s1 general experimental procedure

H3-R40Me1

27KSAPATGGIKKPHR40

y11

y11,2+

y13,2+

y8

y3

y4

y13

y9

y5

y12,2+

b3-H2O

y7

b13

b11-H2O

y6

b1

b3

b9

b11

y2

y12

y10

M/Z: 843.4823, Charge: 2+, Mass Error: -7ppm, Ion Score: 65


Figure s1 general experimental procedure

H3-R49Me1

41FRPGTVALR49

y7

b2

[M-H2O+2H]2+

y8-NH3,2+

[M-2H2O+2H]2+

b7

b6

y8-NH3

y6

y1-NH3

b8

y2-NH3

y3-NH3

b5

b1

y2

y1

y5

M/Z: 551.3031, Charge: 2+, Mass Error: 1ppm, Ion Score: 41


Figure s1 general experimental procedure

H3-K56Me1

53KYQKSTDLLIR63

[M-2H2O+2H]2+

[M-H2O+2H]2+

y8

y4

y9

y3

y6

b3

b10

b8

b2

y9-NH3

y9,2+

b7

y5

y2

b9

y10

M/Z: 754.4066, Charge: 2+, Mass Error: -1ppm, Ion Score: 47


Figure s1 general experimental procedure

H3-K79Me1

73DIAHEFKAELR83

y10,2+

y8,2+

y7

y6

y8

y9,2+

b4

[M-H2O+2H]2+

b9

y5

b6

b10

y2

b3-H2O

y9

b2

y4

y10

M/Z: 727.8854, Charge: 2+, Mass Error: 0ppm, Ion Score: 41


Figure s1 general experimental procedure

H3-R83Me1

73DIAHEFKAELR83

y10,2+

y8,2+

y7

y6

y8

[M-H2O+2H]2+

y9,2+

y4

y5

b4

b7

y3

b5

b8

y9

b6

b2-H2O

b3

b2

y2

M/Z: 727.8854, Charge: 2+, Mass Error: 0ppm, Ion Score: 49


Figure s1 general experimental procedure

H3-R128Me1

116RVTIMTKDMQLAR128

b8

y5

y9

b7

y12

b10

b5

y7

b11

y3-NH3

y8

b4-NH3,2+

b4

y4

b6

y3

b9

M/Z: 844.964, Charge: 2+, Mass Error: 1ppm, Ion Score: 46


Figure s1 general experimental procedure

H3.3-K9AcK14Ac

9KSTGVKAPR17

[M-H2O+2H]2+

[M-2H2O+2H]2+

y8

b7

y3

b1

y2

b6

b5

y8,2+

y7

b2

y4

b3-NH3

b3

y6

y5

b8

M/Z: 549.2891, Charge: 2+, Mass Error: -3ppm, Ion Score: 44


Figure s1 general experimental procedure

H3.3-K9Ac

9KSTGVKAPR17

[M-2H2O+2H]2+

[M-H2O+2H]2+

y8

y4

b7

y3

b6

b1

y2

b5

y7

b2

y6

y1

y8,2+

b3

y5

b8

M/Z: 556.2984, Charge: 2+, Mass Error: 0ppm, Ion Score: 32


Figure s1 general experimental procedure

H3.3-K14Form

9KSTGVKAPR17

[M-H2O+2H]2+

y8

[M-2H2O+2H]2+

b1

y3

b7

y2

b6

y6

b4-H2O

b2

y5

b3-H2O

y7

b8

M/Z: 542.3081, Charge: 2+, Mass Error: -6ppm, Ion Score: 64


Figure s1 general experimental procedure

H3.3-K18AcK23Ac

18KQLATKAAR26

y8

b7

b8

y2

b6-H2O

b4

y3

[M-H2O+2H]2+

y5

b1

y4

y6

y8,2+

y7

y1

b6

b3

b2

b5

M/Z: 563.8318, Charge: 2+, Mass Error: -1ppm, Ion Score: 59


Figure s1 general experimental procedure

H3.3-K23Form

18KQLATKAAR26

y8

b7

b8

y2

b6-H2O

y3

y5

b4

b1

y6

y8,2+

y1

b6

y7

b3

b8-NH3,2+

b2

y4

b5

M/Z: 563.8318, Charge: 2+, Mass Error: -1ppm, Ion Score: 45


Figure s1 general experimental procedure

H3.3-K56Ac

53KYQKTTDLLIR63

y9

y6

y8

b2

b10

y4

b9

y5

b3

y9-NH3

y3

b8

b7

y2

y10

y9,2+

b4

b6

M/Z: 738.9251, Charge: 2+, Mass Error: 1ppm, Ion Score: 56


Figure s1 general experimental procedure

H3.3-K79Me1

73DIAMEMKSDIR83

[M-2H2O+2H]2+

[M-H2O+2H]2+

b7

b8

y5

y2

y4

b3

b4

y7

y3

y8

M/Z: 725.3299, Charge: 2+, Mass Error: 0ppm, Ion Score: 26


Figure s1 general experimental procedure

H4-K4Ac

1AGGKGGK7

y6

[M-2H2O+2H]2+

b5

b6

b4

y3

y4

y5

y2

b3

M/Z: 728.3949, Charge: 2+, Mass Error: 2ppm, Ion Score: 35


Figure s1 general experimental procedure

H4-K4AcK7Ac

1AGGKGGKGMGK11

[M-2H2O+2H]2+

y10

b9

b8

b7

y7

b10

y6

b5

b6

y5

y9

b4

y8

M/Z: 1143.5856, Charge: 1+, Mass Error: 2ppm, Ion Score: 64


Figure s1 general experimental procedure

H4-K7Ac

1AGGKGGKGMGK11

[M-2H2O+2H]2+

b9

[M-H2O+2H]2+

y10,2+

b7

y7

y4

y2

b8

b10

y3

y5

y1

b4

y10

y6

M/Z: 579.3083, Charge: 2+, Mass Error: 2ppm, Ion Score: 71


Figure s1 general experimental procedure

H4-K4AcK7AcK11AcK15Ac

1AGGKGGKGMGKVGAKR16

[M-2H2O+2H]2+

[M-H2O+2H]2+

y15,2+

y4

b12

y5

y7

b11

b14

b15

b9

b10

b7

b13

y2

y3

b5

y8

y10

y11

y12

b4

y9

M/Z: 841.9559, Charge: 2+, Mass Error: 2ppm, Ion Score: 51


Figure s1 general experimental procedure

H4-K4AcK7AcK11Ac

1AGGKGGKGMGKVGAKR16

y15,2+

[M-H2O+2H]2+

y4

b12

y9

b15

b11

y5

y6

b9

b7

y12

b4

b8

b13

y11

b5

b14

y2

y8

y7

b10

M/Z: 848.9645, Charge: 2+, Mass Error: 2ppm, Ion Score: 67


Figure s1 general experimental procedure

H4-K4AcK11Ac

1AGGKGGKGMGKVGAKR16

[M-2H2O+2H]2+

y15,2+

[M-H2O+2H]2+

b12

y4

b11

b15

y5

y12,2+

y8,2+

y9

b14

b7

y7

y2

b6

y13,2+

b9

y14,2+

y9,2+

y3

b5

b13

y8

b10

y12

y11

M/Z: 855.9704, Charge: 2+, Mass Error: 1ppm, Ion Score: 68


Figure s1 general experimental procedure

H4-K7AcK11Ac

1AGGKGGKGMGKVGAKR16

y15,2+

b12

[M-H2O+2H]2+

y4

[M-2H2O+2H]2+

b11

y5

y8,2+

b15

b9

y7

b14

b7

b8

y9

b4

y13,2+

y3

y12

y6

b6

b13

y2

y8

b10

M/Z: 855.9723, Charge: 2+, Mass Error: 3ppm, Ion Score: 62


Figure s1 general experimental procedure

H4-K11Ac

1AGGKGGKGMGKVGAKR16

y15,2+

b12

[M-2H2O+2H]2+

[M-H2O+2H]2+

b15

b11

y4

b8

b9

y5

b7

y3

y12

y8,2+

b14

y8

y9

b4

M/Z: 862.9801, Charge: 2+, Mass Error: 3ppm, Ion Score: 59


Figure s1 general experimental procedure

H4-K7Ub

1AGGKGGKGMGKVGAKR16

y15,2+

y4

b12

y5

b11

b15

y12

b14

y2

b7,2+

b4

b10

y3

b5

b13

b6

y11

M/Z: 870.9785, Charge: 2+, Mass Error: -2ppm, Ion Score: 41


Figure s1 general experimental procedure

H4-K7AcK11AcK15Ac

5GGKGMGKVGAKR16

[M-2H2O+2H]2+

b4

b7

b5

y7-NH3

y9

b6

y11-NH3

y9-NH3

y7

y4

b8

y10

y11

y8

y5-NH3

y5

y6

y3

M/Z: 1327.7186, Charge: 1+, Mass Error: 3ppm, Ion Score: 66


Figure s1 general experimental procedure

H4-K11AcK15Ac

8GMGKVGAKR16

[M-2H2O+2H]2+

b4

b6

b5

y5-NH3

y4-NH3

y7-NH3

y7

y8

y5

y6

b8

b5-NH3

b7

b3

b4-NH3

y8-NH3

y6-NH3

y3

M/Z: 1043.5687, Charge: 1+, Mass Error: 2ppm, Ion Score: 30


Figure s1 general experimental procedure

H4-K15Ac

8GMGKVGAKR16

y4

b8

b5

[M-H2O+2H]2+

y5

y8,2+

b4

y1

y7,2+

b7

y2

y3

y7

y6

b3

b2

M/Z: 529.2958, Charge: 2+, Mass Error: 2ppm, Ion Score: 41


Figure s1 general experimental procedure

H4-R35Me1

24ASIEGITKPAIR35

y8

[M-H2O+2H]2+

y4-NH3

b5

y5-NH3

y8-NH3

y10

y11-NH3

b7-H2O

y9

y6

y4

y5

y11

y6-NH3

y7

M/Z: 1397.761, Charge: 1+, Mass Error: 3ppm, Ion Score: 63


Figure s1 general experimental procedure

H4-R55Me1

46ISSFIYDDSR55

[M-H2O+2H]2+

y5-H2O

y3

b4-H2O

b5

y9-H2O

y7

y6-H2O

y8-H2O

b4

y9

y5

y7-H2O

y8

y6

y4

b5

M/Z: 1285.5776, Charge: 1+, Mass Error: 4ppm, Ion Score: 52


Figure s1 general experimental procedure

H4-R67Me1

56QVLKSFLENVVR67

[M-H2O+2H]2+

b10

b5-H2O

b6-H2O

y4

b4

b7

y8

y4-NH3

y7

y9-NH3

y8-H2O

y10

y11

y6

y9

y5

M/Z: 1575.852, Charge: 1+, Mass Error: 3ppm, Ion Score: 54


Figure s1 general experimental procedure

H4-R77Me1

68DAVTYTEHAR77

y7

y9,2+

y6

[M-H2O+2H]2+

y5

y7,2+

y8,2+

b4

y4

b5-H2O

b9,2+

y3

b3

b2

y5,2+

b2-H2O

b6-H2O

b9

b8

y2

b6

y8

y9

M/Z: 616.7969, Charge: 2+, Mass Error: -3ppm, Ion Score: 43


Figure s1 general experimental procedure

H4-K91Me1

78RKTVTAMDVVYALKR92

y7

b8

[M-H2O+2H]2+

b10

y11

y8

b13

y10

b12

y14

b9

y6

y3-NH3

y5

y9

b2-NH3

b11

y13

y4

b6

y12

y2-NH3

b14

M/Z: 967.0534, Charge: 2+, Mass Error: 3ppm, Ion Score: 76


Figure s1 general experimental procedure

H4-R92Me1

79KTVTAMDVVYALKR92

[M-2H2O+2H]2+

b11

b13

b9

y4

y5

b10

b12

y3

b8

y2

y11

y6

b5-H2O

b6

y13

y9

b10,2+

y8

b11-H2O

b10-H2O

b13-H2O

M/Z: 889.0012, Charge: 2+, Mass Error: 1ppm, Ion Score: 43


Figure s1 general experimental procedure

Figure S4. Down-regulation of H2B C-terminal Phosphorylation

P

HAISEGTKAVTKFSSSTN

H2B-1:

[M-H2O+2H]2+

b10

y13

y15

b3

y8

y14

y17,+2

b11

y7,+2

y10

y11

y13,+2

b3-H2O

y16

y17

M/Z:1028.9874, Charge: 2+, ΔM: 1ppm, Ion score: 38


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