Dataset Information

Fishing for newly synthesized proteins with phosphonate-handles

ABSTRACT: Bioorthogonal chemistry introduces affinity-labels into biomolecules with minimal disruption to the original system and is widely applicable in a range of contexts. In proteomics, immobilized metal affinity chromatography (IMAC) enables enrichment of phosphopeptides with extreme sensitivity and selectivity. Here, we adapt and combine these superb assets in a new enrichment strategy using phosphonate-handles, which we term ‘PhosID’. In this approach, ‘click-able’ phosphonate-handles are introduced into proteins via 1,3-dipolar Huisgen-cycloaddition to azido-homo-alanine (AHA) and IMAC is then used to enrich exclusively for phosphonate-labeled peptides. In interferon-gamma (IFNγ) stimulated cells, PhosID enabled the identification of a large number of IFN responsive newly synthesized proteins (NSPs) whereby we monitored the differential synthesis of these proteins over time. Collectively, these data validate the excellent performance of PhosID with efficient analysis and quantification of hundreds of NSPs by single LC-MS/MS runs. We envision PhosID as an attractive and alternative tool for studying stimuli-sensitive proteome subsets.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Jurkat Cell, Hela Cell

SUBMITTER: wei wu

LAB HEAD: Wei Wu

PROVIDER: PXD018272 | Pride | 2020-07-28

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20190607_F1_Ag5_Klein067_SA_FK190509_FT_sample_1_BSA.raw	Raw
	20190607_F1_Ag5_Klein067_SA_FK190509_FT_sample_2_BSA-azide.raw	Raw
	20190607_F1_Ag5_Klein067_SA_FK190509_FT_sample_3_BSA-azide_Palkyne.raw	Raw
	20190607_F1_Ag5_Klein067_SA_FK190509_FT_sample_4_BSA-azide_CuAAC.raw	Raw
	20190607_F1_Ag5_Klein067_SA_FK190509_FT_sample_5_BSA_Pdbco.raw	Raw

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Publications

Cutting blade dentitions in squaliform sharks form by modification of inherited alternate tooth ordering patterns.

Underwood Charlie C Johanson Zerina Z Smith Moya Meredith MM

Royal Society open science 20161130 11

The squaliform sharks represent one of the most speciose shark clades. Many adult squaliforms have blade-like teeth, either on both jaws or restricted to the lower jaw, forming a continuous, serrated blade along the jaw margin. These teeth are replaced as a single unit and successor teeth lack the alternate arrangement present in other elasmobranchs. Micro-CT scans of embryos of squaliforms and a related outgroup (Pristiophoridae) revealed that the squaliform dentition pattern represents a highl ...[more]

PMID: 28018617

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