Dataset Information

Optimization of protocols for detection of de novo protein synthesis in whole blood samples via azide-alkyne cycloaddition

ABSTRACT: Aberrant protein synthesis and protein expression are a hallmark of many disorders ranging from cancer Aberrant protein synthesis and protein expression are a hallmark of many disorders ranging from cancer to neuropsychiatry. Blood-based biomarkers indicative of changes in proteomes have long been held to be potentially useful with respect to disease prognosis and treatment. However, most effort has been focused on unlabelled plasma proteomics which includes non-myeloid origin proteins and no method of dynamically tagging acute changes in proteomes. Herein we report a method for evaluating de novo protein synthesis in whole blood liquid biopsies. Using a modification of the “bioorthogonal noncanonical amino acid tagging” (BONCAT) protocol, rodent whole blood samples were incubated with L-azidohomoalanine (AHA) to allow incorporation of this selectively reactive non-natural amino acid within nascent polypeptides. Overall, we present a rapid and convenient de novo protein synthesis assay usable with whole blood biopsies that can quantify translational change as well as identity of proteins differentially expressed that may be useful for clinical applications

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Rattus Norvegicus (rat)

TISSUE(S): Blood Cell, Blood

SUBMITTER: Janani Priya Venkatasubramani

LAB HEAD: Dr. Aditi Bhattacharya

PROVIDER: PXD019243 | Pride | 2020-10-08

REPOSITORIES: Pride

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Optimization of Protocols for Detection of De Novo Protein Synthesis in Whole Blood Samples via Azide-Alkyne Cycloaddition.

Bowling Heather L HL Kasper Amanda A Patole Chhaya C Venkatasubramani Janani Priya JP Leventer Sarah Parker SP Carmody Erin E Sharp Kevin K Berry-Kravis Elizabeth E Kirshenbaum Kent K Klann Eric E Bhattacharya Aditi A

Journal of proteome research 20200820 9

Aberrant protein synthesis and protein expression are a hallmark of many conditions ranging from cancer to Alzheimer's. Blood-based biomarkers indicative of changes in proteomes have long been held to be potentially useful with respect to disease prognosis and treatment. However, most biomarker efforts have focused on unlabeled plasma proteomics that include nonmyeloid origin proteins with no attempt to dynamically tag acute changes in proteomes. Herein we report a method for evaluating de novo ...[more]

PMID: 32786687

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