Evaluation of female Aedes aegypti proteome via LC-ESI-MS/MS using two protein extraction methods

Abubakar Shettima; Intan H Ishak; Syahirah Hanisah Abdul Rais; Hadura Abu Hasan; Nurulhasanah Othman

doi:10.7717/peerj.10863

Evaluation of female Aedes aegypti proteome via LC-ESI-MS/MS using two protein extraction methods

PeerJ. 2021 Mar 3:9:e10863. doi: 10.7717/peerj.10863. eCollection 2021.

Authors

Abubakar Shettima^{1

2}, Intan H Ishak^{3

4}, Syahirah Hanisah Abdul Rais¹, Hadura Abu Hasan^{3

4}, Nurulhasanah Othman¹

Affiliations

¹ Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Gelugor, Pulau Pinang, Malaysia.
² Department of Microbiology, Faculty of Science, University of Maiduguri, Maiduguri, Borno State, Nigeria.
³ School of Biological Sciences, Universiti Sains Malaysia, Gelugor, Pulau Pinang, Malaysia.
⁴ Vector Control Research Unit (VCRU), School of Biological Sciences, Universiti Sains Malaysia, Gelugor, Pulau Pinang, Malaysia.

Abstract

Background: Proteomic analyses have broadened the horizons of vector control measures by identifying proteins associated with different biological and physiological processes and give further insight into the mosquitoes' biology, mechanism of insecticide resistance and pathogens-mosquitoes interaction. Female Ae. aegypti ingests human blood to acquire the requisite nutrients to make eggs. During blood ingestion, female mosquitoes transmit different pathogens. Therefore, this study aimed to determine the best protein extraction method for mass spectrometry analysis which will allow a better proteome profiling for female mosquitoes.

Methods: In this present study, two protein extractions methods were performed to analyze female Ae. aegyti proteome, via TCA acetone precipitation extraction method and a commercial protein extraction reagent CytoBuster^TM. Then, protein identification was performed by LC-ESI-MS/MS and followed by functional protein annotation analysis.

Results: The CytoBuster^TM reagent gave the highest protein yield with a mean of 475.90 µg compared to TCA acetone precipitation extraction showed 283.15 µg mean of protein. LC-ESI-MS/MS identified 1,290 and 890 proteins from the CytoBuster^TM reagent and TCA acetone precipitation, respectively. When comparing the protein class categories in both methods, there were three additional categories for proteins identified using CytoBuster^TM reagent. The proteins were related to scaffold/adaptor protein (PC00226), protein binding activity modulator (PC00095) and intercellular signal molecule (PC00207). In conclusion, the CytoBuster^TM protein extraction reagent showed a better performance for the extraction of proteins in term of the protein yield, proteome coverage and extraction speed.

Keywords: LC-ESI-MS/MS; Protein extraction methods; Protein identification; Female Aedes aegypti; Functional analysis.

Grants and funding

This work was supported by Universiti Sains Malaysia, Research University Individual Grant (RUI) CIPPM.1001.8012228. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.