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Table 1 Elution time and parent ion masses of candidate glycerophosphocholine species bound to proteins in dialyzed α-synuclein KO cytosolic extracts identified by LC-ESI-MS

From: RETRACTED ARTICLE: Differential regulation of wild-type and mutant alpha-synuclein binding to synaptic membranes by cytosolic factors

Parent ion mass (± 0.15 m/z) a

LC elution time (± 0.4 min) a

Lipid species in complex with proteins in KO cytosol b

Species abundance relative to Wt (Fold change)

Species bound to α-syn following immunoprecipitation (Fold change above non-specific binding) c

494.7

12.29

C14:1-PAF

↑-fold

 
 

12.89

C16:1-LPC

No change

 

496.8

13.79

C14:0-PAF

↑3-fold

↑9.2x

 

14.39

C16:0 LPC

↑2-fold

↑9.8y

520.7

13

C16:2-PAF

↑2-fold

 
 

13.41

C18:2-LPC

No change

 

522.8

14.66

C16:1-PAF

↑5-fold

 
 

15.15

C18:1-LPC

↑2-fold

 

524.9

17.2

C16:0-PAFc

↑2-fold

↑1.7x

 

18.11

C18:0-LPC

↑4-fold

↑2.3y

545

 

C18:4-PAF

↑11-fold

 
 

13.12

C20:4-LPC

  

545.9

13.11

C18:3-PAF

De novo detection

 
 

13.99

C20:3-LPC

  

568.8

13.12

C20:6-PAF

↑4-fold

 
  

C22:6-LPC

  

581

15.32

C20:0-PAF

↑17-fold

 
 

16.04

C22:0-LPC

↑12-fold

 

594

16.52

C24:7-LPC

↑2-fold

 
  

C22:7-PAF

  
  

23:7c

  
  

24:7d

  
  

C20:0-acyl-PAF

  
  

C24:0-lysoPAF

  
  1. aVariations between m/z and retention time between runs were established for all glycerophospholipid species and respresents mean ± standard deviation.
  2. bIdentification is predicted based on the theoretical monoisotopic mass values. CX:Y refers to the number of carbon atoms and double bonds in the sn-1 chain with a predicted acetyl (PAF) or hydroxyl (LPC) group at the sn-2 position. Only the most likely species are indicated although multiple isoforms may be present with the double bond in the alkyl chain at different positions. Isobaric species with same m/z eluting at different times were not further distinguished with the exception of C16:0 PAF.
  3. cReplicate experiments were performed in which α-syn was immunoprecipitated from Wt cytosolxor recombinant α-syn was added to KO cytosoly. Immunoprecipitates were analysed by LC-ESI-MS. Data represent mean increase in relative abundance above background (non-specific) signal ± standard deviation as described in Materials and Methods.
  4. dIdentity verified by based on its coelution with d4-C16-PAF spiked at time of analysis.