Mass spectrometry identification of age-associated proteins from the malaria mosquitoes Anopheles gambiae s.s. and Anopheles stephensi.

James Monkman,Marcus L. Hastie,Roger L. Kitching,Gerry F. Killeen,Leon E. Hugo,Keyur A. Dave,Patricia E. Dale,Maggy T. Sikulu,Jeffry J. Gorman,Brian H. Kay

doi:10.1016/j.dib.2015.07.007

Abstract

This study investigated proteomic changes occurring in Anopheles gambiae and Anopheles stephensi during adult mosquito aging. These changes were evaluated using two-dimensional difference gel electrophoresis (2D-DIGE) and the identities of aging related proteins were determined using capillary high-pressure liquid chromatography (capHPLC) coupled with a linear ion-trap (LTQ)-Orbitrap XL hybrid mass spectrometry (MS). Here, we have described the techniques used to determine age associated proteomic changes occurring in heads and thoraces across three age groups; 1, 9 and 17 d old A. gambiae and 4 age groups; 1, 9, 17 and 34 d old A. stephensi. We have provided normalised spot volume raw data for all protein spots that were visible on 2D-DIGE images for both species and processed Orbitrap mass spectrometry data. For public access, mass spectrometry raw data are available via ProteomeXchange with identifier PXD002153. A detailed description of this study has been described elsewhere [1].

Highlights

Mass spectrometry identification of ageassociated proteins from the malaria mosquitoes Anopheles gambiae s.s. and Anopheles stephensi
This study investigated proteomic changes occurring in Anopheles gambiae and Anopheles stephensi during adult mosquito aging
We have described the techniques used to determine age associated proteomic changes occurring in heads and thoraces across three age groups; 1, 9 and 17 d old A. gambiae and 4 age groups; 1, 9, 17 and 34 d old A. stephensi

Summary

Mosquito preparation and protein extraction

Two cohorts each of A. gambiae and A. stephensi were reared at different times. For each cohort, adults were collected 24 h post emergence and again at 9 and 17 d post emergence for A. gambiae and at 9, 17 and 34 d post emergence for A. stephensi. Samples were resuspended in 2D buffer and total protein content was quantified using the 2D quantification kit (GE Healthcare) following the manufacturer's protocol. Each sample pool was combined with 4.5 ml of 100 Â ampholytes (Bio-Rad, Richmond, CA, USA.), 5.6 ml Destreak reagent (GE Healthcare) and made up to 450 ml in 2D Buffer. The strips were transferred to a new focussing tray, overlayed with mineral oil and allowed to rehydrate overnight at room temperature. The strips were overlayed with fresh mineral oil and iso-electric focussing was performed using the following run conditions: 50 μA per strip, 250 V for 15 min, 1000 V for 5 h, 10,000 V for 4 h and 500 V to reach a total of 80,000 V hÀ1. Electrophoresis was performed in a Proteans Plus Dodeca cell (Bio-Rad) at 5 mA/gel for 15 min, 10 mA/gel for 15 min and 30 mA/gel until the dye front reached the bottom of the gel

Imaging and processing

Sample preparation for MS

Findings

Data analysis