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Biochemical Studies of Recombinant AaCHYMO from the Female Aedes Aegypti Mosquito
Author(s) -
Burata Olive,
Rascon Alberto A.
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.918.9
Subject(s) - aedes aegypti , blood meal , biology , midgut , proteases , digestion (alchemy) , population , aedes , dengue fever , zoology , enzyme , biochemistry , immunology , ecology , chemistry , medicine , larva , environmental health , chromatography
The new emergence of the Zika arbovirus within the Americas has brought great attention to the female Aedes aegypti mosquito. This anthropophilic mosquito undergoes multiple blood feedings throughout its lifetime in order to successfully provide nutrients for the gonotrophic cycle, the process of egg laying and oocyte maturation. In doing so, the Ae. aegypti has conveniently become the dominant vector in transmitting Zika and various other arboviruses. Zika virus infection has already been linked to microcephaly in newborn babies, an abnormal development of the brain characterized by a significantly smaller than average head size. Current defenses against these mosquitos mainly involve an exaggerated usage of pesticides that have led to an increase in pesticide‐resistant mosquitos, as well as collateral injury towards pollinators. Since midgut serine proteases within the Ae. aegypti are required for digestion of blood meal proteins into the nutrients required to fuel the gonotrophic cycle, inhibiting these enzymes may provide a more specific method of population control. Blood meal digestion has been shown to have an early and a late phase, with the majority of blood meal digestion and trypsin‐like activity being observed in the late phase. However, little is known about a different class of serine proteases, the chymotrypsin‐like serine proteases. One chymotrypsin‐like enzyme, AaCHYMO, was previously reported to be expressed in the early phase of midgut digestion, but no biochemical information is available and its role in the blood meal digestion process is unknown. Therefore, a recombinant form of AaCHYMO was constructed using the bacterial expression pET28a vector and expressed using Shuffle T7 E. coli cells. Purification of the recombinant form of AaCHYMO was successful, but surprisingly the enzyme isolated was the active form, potentially indicating the possibility of this chymotrypsin to be auto‐catalytic. Support or Funding Information SC3 Research Grant from NIH