Dataset Information

Lysine Methylation Promotes VEGFR-2 Activation and Angiogenesis

ABSTRACT: We have used a chimeric VEGFR-2 in which the extracellular domain of mouse VEGFR-2 was replaced with the extracellular domain of human CSF-1 receptor. VEGFR-2 was immunoprecipitated with anti-VEGFR-2 antibody from PAE cells ectopically expressing VEGFR-2. The immunoprecipitated proteins were eluted and separated on SDS-PAGE, followed by in-gel chymotrypsin or trypsin digestion. The digested samples were analyzed by nano LC/MS/MS on a Thermo Fisher LTQ Orbitrap XL. The LC-MS/MS data were analyzed using Proteome Discoverer (Thermo Fisher Scientific; Version 1.3.0.339). MS/MS search was carried out using Sequest search algorithm against the sequence of target mouse protein from the UniProtKB database. Search parameters included chymotrypsin as the enzyme with four missed cleavage allowed; methylation at lysine and arginine, phosphorylation of serine, threonine, and tyrosine, alkylation at cysteine, and oxidation of methionine were set as dynamic modifications. Precursor and fragment mass tolerance were set to 5 ppm and 0.8 Da, respectively. The false discovery rate was calculated by enabling the peptide sequence analysis using a decoy database. High confidence peptide identifications were obtained by setting a target false discovery rate threshold of 1% at the peptide level.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

TISSUE(S): Huvec Cell

SUBMITTER: Yan Jiang

LAB HEAD: Nader Rahimi

PROVIDER: PXD000520 | Pride | 2019-11-22

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	WT_chymotrypsin_ingel.RAW	Raw
	WT_chymotrypsin_ingel_Sequest.msf	Msf
	WT_chymotrypsin_ingel_Sequest.pep.xml	Pepxml

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Publications

Lysine methylation promotes VEGFR-2 activation and angiogenesis.

Hartsough Edward J EJ Meyer Rosana D RD Chitalia Vipul V Jiang Yan Y Marquez Victor E VE Zhdanova Irina V IV Weinberg Janice J Costello Catherine E CE Rahimi Nader N

Science signaling 20131203 304

Activation of vascular endothelial growth factor receptor-2 (VEGFR-2), an endothelial cell receptor tyrosine kinase, promotes tumor angiogenesis and ocular neovascularization. We report the methylation of VEGFR-2 at multiple Lys and Arg residues, including Lys(1041), a residue that is proximal to the activation loop of the kinase domain. Methylation of VEGFR-2 was independent of ligand binding and was not regulated by ligand stimulation. Methylation of Lys(1041) enhanced tyrosine phosphorylation ...[more]

PMID: 24300896

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