Dataset Information

Mosquito-Derived Anophelin Sulfoproteins are Potent Anti-Thrombotics

ABSTRACT: we first expressed AAa and AAg in a eukaryotic system to investigate whether the proteins could be sulfated in vivo. For this purpose we used insect cells as a model of the Anopheles mosquito. Specifically, codon-optimized sequences encoding AAa and AAg were designed as N-terminal fusions with the honeybee mellitin signal sequence in order to direct the recombinant proteins to the secretion pathway and were expressed in Trichoplusia ni insect cells. Following expression, the cell medium containing the secreted proteins was analyzed by nanoliquid chromatography coupled to tandem mass spectrometry (nanoLC-MS/MS).

INSTRUMENT(S): Orbitrap Fusion ETD

ORGANISM(S): Anopheles Albimanus Anopheles Gambiae (african Malaria Mosquito)

SUBMITTER: Benjamin Parker

LAB HEAD: Benjamin Parker

PROVIDER: PXD008240 | Pride | 2018-08-30

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20170609_Fusion_BP_RichPayne_Baclo_Anophelin_Digest.raw	Raw
	20170609_Fusion_BP_RichPayne_Baclo_Anophelin_Digest_2.raw	Raw
	20170609_Fusion_BP_RichPayne_Baclo_Anophelin_Intact.raw	Raw
	20170609_Fusion_BP_RichPayne_Baclo_Anophelin_Intact_2.raw	Raw
	20170814_Fusion_BP_ETD_RichPayne_AnophelinSynthetic_Y12_300fmol_1.raw	Raw

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Publications

Mosquito-Derived Anophelin Sulfoproteins Are Potent Antithrombotics.

Watson Emma E EE Liu Xuyu X Thompson Robert E RE Ripoll-Rozada Jorge J Wu Mike M Alwis Imala I Gori Alessandro A Loh Choy-Theng CT Parker Benjamin L BL Otting Gottfried G Jackson Shaun S Pereira Pedro José Barbosa PJB Payne Richard J RJ

ACS central science 20180328 4

The anophelins are small protein thrombin inhibitors that are produced in the salivary glands of the Anopheles mosquito to fulfill a vital role in blood feeding. A bioinformatic analysis of anophelin sequences revealed the presence of conserved tyrosine residues in an acidic environment that were predicted to be post-translationally sulfated in vivo. To test this prediction, insect cell expression of two anophelin proteins, from Anopheles albimanus and Anopheles gambiae</ ...[more]

PMID: 29721529

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Project description:Neurodevelopment is a tightly coordinated process, in which genome is exposed to spectra of endogenous agents at different stages of differentiation. Increasing body of evidence suggests that DNA damage is an important feature of developing brain, tightly linked to gene expression programs and neuronal activity. Some of the most frequent DNA damage includes changes to DNA bases, recognized by DNA glycosylases and repaired through base excision repair (BER). The only mammalian DNA glycosylase able to remove frequent alkylated DNA based is alkyladenine DNA glycosylase (Aag, aka Mpg). Recently we showed that, besides participation in DNA repair, AAG affects expression of neurodevelopmental genes in human cells. Aag was further proposed to act as reader of epigenetic marks, including 5-hydroxymethylcytosine (5hmC), in the mouse brain. Despite the evidence of potential Aag involvement in the key brain processes, the impact of Aag loss on developing brain remains unknown. Here, by using Aag knockout (Aag-/-) mice, we show that Aag absence leads to reduced DNA damage levels, evident in lowered number of γH2AX foci in P5 hippocampi. This is accompanied by changes in 5hmC signal intensity in different hippocampal regions. Analysis of gene expression in hippocampus and prefrontal cortex, at multiple developmental stages, indicates that lack of Aag results in altered gene expression, primarily of genes involved in regulation of response to stress. One of the most prominent genes deregulated in Aag-dependent manner, at all tested developmental stages, is aldehyde dehydrogenase 2 (Aldh2). In line with the changes in hippocampal DNA damage levels and the gene expression, adult Aag-/- mice exhibit altered behavior, evident in decreased anxiety in the Elevated Zero Maze and increased alternations in the Elevated T Maze tests. Taken together these results suggests that Aag has functions in modulation of genome dynamics during brain development, important for animal behavior.