Table I.

H1 phosphorylation during interphase and mitosis in HeLa S3 cells

HIC peak Molecular mass Δm Variant Modifications 
 Measured Predicted    
 D D    
Interphase 
21,491.6 21,332.8 +160 H1.4 pS172, pS187 
21,412.3 21,332.8 +80 H1.4 pS187 
21,333.1 21,332.8 H1.4 None 
20,912.1 20,832.6 +80 H1.2 (A142) pS173 
2,0941.7 20,832.6 +110 H1.2 (T142) pS173 
20,832.3 20,832.6 H1.2 (A142) None 
20,862.2 20,832.6 +30 H1.2 (T142) None 
Mitosis 
6p-H1.4 21,813.9 21,332.8 +480 H1.4 pT18, pS27, pT146, pT154, pS172, pS187 
4p-H1.2 21,153.3 20,832.6 +320 H1.2 (A142) pT31, pT146, pT154, pS173 
4p-H1.2 21,182.9 20,832.6 +350 H1.2 (T142) pT31, pT146, pT154, pS173 
HIC peak Molecular mass Δm Variant Modifications 
 Measured Predicted    
 D D    
Interphase 
21,491.6 21,332.8 +160 H1.4 pS172, pS187 
21,412.3 21,332.8 +80 H1.4 pS187 
21,333.1 21,332.8 H1.4 None 
20,912.1 20,832.6 +80 H1.2 (A142) pS173 
2,0941.7 20,832.6 +110 H1.2 (T142) pS173 
20,832.3 20,832.6 H1.2 (A142) None 
20,862.2 20,832.6 +30 H1.2 (T142) None 
Mitosis 
6p-H1.4 21,813.9 21,332.8 +480 H1.4 pT18, pS27, pT146, pT154, pS172, pS187 
4p-H1.2 21,153.3 20,832.6 +320 H1.2 (A142) pT31, pT146, pT154, pS173 
4p-H1.2 21,182.9 20,832.6 +350 H1.2 (T142) pT31, pT146, pT154, pS173 

HIC peaks are labeled as in Fig. 1 B. Molecular masses are reported as neutral monoisotopic species. Predicted values were found using NCBI Protein database accession no. NP_005312 (H1.4) and NP_005310 (H1.2), assuming the loss of Met1 during protein maturation in vivo and residues 2–5 during electrospray ionization. Peak identifications are based on MS/MS sequencing of multiple electron capture dissociation fragment ions. The A142 and T142 allelic variants of H1.2 were detected initially by MS and confirmed by genotyping as described in Fig. S1. Phosphorylated residues were identified by MS/MS sequencing of electron capture dissociation fragment ions. Analyses of the H1.2 (T142) forms indicated that phosphorylated residues were identical to those determined for the H1.2 (A142) forms.

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