XB-ART-51525J Biochem April 1, 2016; 159 (4): 407-19.
Exploring the phosphoproteome profiles during Xenopus egg activation by calcium stimulation using a fully automated phosphopeptide purification system.
To explore the phosphoproteome profiles duringXenopusegg activation by Ca(2+)-stimulation, an automated phosphopeptide purification system involving a titania column was improved by introducing 4-step elution with phosphate buffers. The number of detected phosphopeptides in the tryptic digest of aXenopusegg cytosol fraction on mass spectrometry (MS) was increased 1.5-fold and the percentage of multiply phosphorylated peptides increased from 17 to 24% with introduction of the 4-step elution method. Phosphopeptides were purified by the improved method from tryptic digests of cytosol fractions ofXenopuseggs without and with a Ca(2+)-stimulus, and then, analysed by MS. One thousand three hundred and seventy-five and 994 phosphopeptides were reproducibly detected on duplicate MS, respectively. They included 818 and 437 phosphopeptides specific to each digest, respectively. A method involving isobaric tags for relative and absolute quantitation (iTRAQ) was also applied to compare the phosphorylation levels inXenopuseggs without and with a Ca(2+)-stimulus, the ratios for 112 phosphopeptides in tryptic digests of these egg cytosol fractions being obtained. It was suggested from all the results that the phosphorylation sites and levels change duringXenopusegg activation for many known and unknown sites on structural proteins, signalling related proteins, cell cycle-related proteins and others.
PubMed ID: 26530081
PMC ID: PMC4885929
Article link: J Biochem
Species referenced: Xenopus laevis
Genes referenced: abcf1 akt2 cdk11b cfl1 ctps1 dbnl eif4ebp2 ldlrap1 lmnb3 mapk1 mcm2 melk nap1l1 ncam1 ndrg2 nsfl1c pnpo rnaseh2b rps6ka3 septin2 sf1 siva1 stmn1 tfdp1 ttk usp8
GO keywords: phosphorylation
References [+] :
Backs, The gamma isoform of CaM kinase II controls mouse egg activation by regulating cell cycle resumption. 2010, Pubmed