Sequence

Rat hepatocytes

Mouse Brown Adipocytes

3T3-L1

Site  -5 -4 -3 -2 -1 0 +1 +2 +3 +4 +5

None [4]

Ins [3]

AICAR [3]

cAMP [3]

Iso [1]

None [1]

Ins [2]

Fors [1]

CL [1]

None [1]

Ins [1]

cAMP [1]

di8 [1]

M1                             M  E   V    H    E     L  

xxxx

xxx

xxx

xxx

x

xx

x

x

x

x

x

x

x

K49    Y   A  T    R   P  K  A    L    K    P     P

x

 

 

 

 

 

 

 

 

 

 

 

 

K52    R  P  K    A   L  K   P    P    C    D    L

x

x

x

xx

x

 

x

x

 

x

 

 

x

K217  E  G  V    E   N  K  L    T    P    C     L

xxxx

xx

xxx

xxx

 

xx

x

x

x

x

x

 

 

K223  L  T   P    C  L   K  I     I    V    I      M

xxxx

xx

xxx

xxx

x

xx

x

x

x

x

x

 

 

K341  L  C  H    G   A  K  I     G   F    F     Q

 

 

 

 

x

 

 

 

 

 

 

 

 

K387  A  N  T    S   V  K  R    W   L    L     D

 

 

x

x

 

 

 

 

 

 

 

 

 

K504  N  Y  Q    A   A  K  G    E   G    E     V

 

 

x

 

 

 

 

 

 

 

 

 

 

K533  A  E  A    L   D  K  D    G   W    L     H

 

x

 

 

 

 

 

 

 

 

 

 

 

K544  T  G  D    I   G   K W    L   P    N     G

 

x

 

 

 

 

 

 

 

 

 

 

 

K562  K  K  H   I    F   K  L    A    Q    G    E

 

xxx

xx

x

x

x

 

 

x

x

 

 

x

x, Identified acetylated lysines. Each “x” indicates that the modification was identified and multiple “x’s” indicate that the modification was identified by the corresponding number of replicates. For example, “xx” indicates that the modification was identified in 2 replicates.
[ ] Number of replicates.
Abbreviations: Ins, insulin; AICAR, 5-aminoimidazole-4-carboxamide riboside; cAMP, dibutyryl-cyclic adenosine monophosphate; Iso, isoproterenol; Fors, forskolin; CL, CL-316243; di8,1,2-dioctanoyl-sn-glycerol
Note: ACSL1 was immunopurified from samples using RIPA buffer. Some of the untreated liver and brown adipocyte immunopurifications were done with gentle lysis buffer
Table 1: Acetylation sites found in overexpressed ACSL1. Ad-Acsl1-Flag was overexpressed in rat hepatocytes, mouse brown adipocytes or 3T3-L1 adipocytes, and treated as indicated.