SILAC-Based Quantitative Proteomic Analysis of Drosophila Embryos.

Methods Mol Biol

Developmental Genetics Group, Institute for Biology, University of Kassel, Kassel, Germany.

Published: November 2022

The fruit fly Drosophila melanogaster represents a classic genetic model organism that is amenable to a plethora of comprehensive analyses including proteomics. SILAC-based quantitative proteomics is a powerful method to investigate the translational and posttranslational regulation ongoing in cells, tissues, organs, and whole organisms. Here we describe a protocol for routine SILAC labeling of Drosophila adults within one generation to produce embryos with a labeling efficiency of over 92%. In combination with genetic selection markers, this method permits the quantification of translational and posttranslational changes in embryos mutant for developmental and disease-related genes.

Download full-text PDF

Source
http://dx.doi.org/10.1007/978-1-0716-2863-8_15DOI Listing

Publication Analysis

Top Keywords

silac-based quantitative
8
translational posttranslational
8
quantitative proteomic
4
proteomic analysis
4
analysis drosophila
4
drosophila embryos
4
embryos fruit
4
fruit fly
4
fly drosophila
4
drosophila melanogaster
4

Similar Publications

SILAC-Based Characterization of Plasma-Derived Extracellular Vesicles in Patients Undergoing Partial Hepatectomy.

Int J Mol Sci

October 2024

Department of Vascular Biology and Thrombosis Research, Centre of Physiology and Pharmacology, Medical University of Vienna, Schwarzspanierstrasse 17, A-1090 Vienna, Austria.

Article Synopsis
  • Post-hepatectomy liver failure (PHLF) poses a major risk for patients after partial liver surgery, with a need for better prognostic markers and treatments to boost liver recovery.
  • A study analyzed plasma samples from nine hepatocellular carcinoma (HCC) patients before and after surgery to identify proteins in extracellular vesicles (EVs) that might indicate liver regeneration status.
  • The findings revealed significant changes in protein levels in patients with PHLF, highlighting disruptions in critical cellular processes and suggesting potential biomarkers that were present before the surgery.
View Article and Find Full Text PDF

p53 deficiency mediates cisplatin resistance by upregulating RRM2 and crotonylation of RRM2 through the downregulation of SIRT7.

Front Mol Biosci

June 2024

Central Laboratory, Peking University School and Hospital of Stomatology, National Center for Stomatology, National Clinical Research Center for Oral Diseases, National Engineering Research Center of Oral Biomaterials and Digital Medical Devices, Beijing Key Laboratory of Digital Stomatology, NHC Key Laboratory of Digital Stomatology, NMPA Key Laboratory for Dental Materials, Beijing, China.

p53 deficiency plays a crucial role in chemotherapy resistance through various biological events, including posttranslational modifications (PTMs). Recently, lysine crotonylation (Kcr) has been shown to play a vital role in cancer progression. However, the global p53-regulated crotonylome and the function of these altered Kcr proteins after p53 deficiency remain unclear.

View Article and Find Full Text PDF

The expression level of individual proteins varies markedly during the progression of the growth phase in bacteria. A set of proteins was quantified in total proteome during 14 days of batch cultivation using pulse stable isotope labeled amino acids in cell culture (SILAC)-based quantitative mass spectrometry.

View Article and Find Full Text PDF

A SILAC-based accurate quantification of shrimp allergen tropomyosin in complex food matrices using UPLC-MS/MS.

Food Chem

May 2024

National Key Laboratory of Veterinary Public Health and Safety, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China; Beijing Key Laboratory of Diagnostic and Traceability Technologies for Food Poisoning, Beijing Center for Disease Control and Prevention, Beijing 100013, China; School of Public Health, Capital Medical University, Beijing 100069, China. Electronic address:

The carryover of trace allergens in complex food matrices poses challenges for detection techniques. Here, we demonstrate an accurate UPLC-MS/MS quantification assay for the shrimp allergen tropomyosin with a full-length isotope-labelled recombinant tropomyosin (TM-I) internal standard in complex food matrices. The TM-I, expressed based on the SILAC technique, exhibited a high isotope labelling ratio (>99%), purity, and alignment with the natural sequence.

View Article and Find Full Text PDF

SILAC proteomics based on 3D cell spheroids unveils the role of RAC2 in regulating the crosstalk between triple-negative breast cancer cells and tumor-associated macrophages.

Int J Biol Macromol

January 2024

Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, PR China; Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education, Department of Integration of Chinese and Western Medicine, Peking University Cancer Hospital and Institute, Beijing 100142, PR China. Electronic address:

Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer, and is characterized by a high infiltration of tumor-associated macrophages (TAMs). TAMs contribute significantly to tumor progression by intricately interacting with tumor cells. Deeply investigating the interaction between TNBC cells and TAMs is of great importance for finding potential biomarkers and developing novel therapeutic strategies to further improve the clinical outcomes of TNBC patients.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!