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Table 1 Proteomic methods to enrich and detect post-translational modifications

From: Functional decorations: post-translational modifications and heart disease delineated by targeted proteomics

Modification

Amino acid

Common motifs

Enrichment methods

Proteomic technologies

Phosphorylation

S, T, Y

>320 possibilities

Radiolabeling

Immobilized metal affinity chromatography

TiO2 enrichment

IP

2-DE, MS/MS

LC-MS/MS

LC-MS/MS

2-DE, LC-MS/MS

N-glycosylation

N

N-x-S/T/C

Glycan staining

Lectin affinity

Interaction chemistry

2-DE, MS/MS

2-DE, MS/MS

LC-MS/MS

O-GlcNAcylation

S, T

P-V-S/T

Chemical tagging

Affinity isolation

IP

LC-MS/MS

LC-MS/MS

2-DE, LC-MS/MS

Redox

C, M, Y, W,

C/S/T-x-x-C,

C-x-x-C/S/T

Biotin-switch assay

Thiol disulphide exchange

Fluorescent labeling

2-DE, LC-MS/MS

LC-MS/MS

2-DE, MS/MS

Advanced glycation end products

  

Boronate affinity chromatography

IP

Western blotting

LC-MS/MS

2-DE, MS/MS

SDS-PAGE

Proteolytic cleavage

S, C, D, K, R

Specific to each protease

In vitro assays

TAILS

2-DE, LC-MS/MS

LC-MS/MS

Deamidation

N, Q

N-G, N-S

 

Gel electrophoresis

MS

Sumoylation

K

Ψ-K-x-D/E

IP

Enzymatic reaction

2-DE, LC-MS/MS

LC-MS/MS

SDS-PAGE

Citrullination

R

G-x-R-G-Ψ

Chemical derivitization

 

Methylation

R, K, Q

M-K, R-G-G, R-G-X, R-X-G, W-x-x-x-R

Western blotting

IP

SDS-PAGE

2-DE, LC-MS/MS

Lysine acetylation

K

G-K, K-P

IP

2-DE, LC-MS/MS

  1. Ψ indicates a hydrophobic residue. 2-DE, two-dimensional gel electrophoresis; IP, immunoprecipitation; LC, liquid chromatography; MS/MS, tandem mass spectroscopy; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; TAILS, terminal amine isotopic labeling of substrates.