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Expression of Aedes aegypti painless protein for structural studies : Production, solubilization, and purification of membrane proteins

Farkas, Titus LU (2024) KEMR30 20242
Department of Chemistry
Abstract
Aedes aegypti is one of the most lethal disease vectors in modern history, spreading a majority of the deadliest diseases mediated by mosquitos. To try and combat this, mankind has, for a long time, tried to create repellents to keep the insects away. These repellents affect receptors in the mosquitos, keeping them at bay. One receptor that is highly expressed in the mosquito’s prey/host sensory organ is the Painless protein. By better understanding this receptor, more effective repellents might be available. This project delves into the structural studies of the protein by expressing it in Pichia pastoris and purifying it using a His tag and an attached GFP tag. The aim is to use Cryo-EM to determine its structure. Electrophysiology was... (More)
Aedes aegypti is one of the most lethal disease vectors in modern history, spreading a majority of the deadliest diseases mediated by mosquitos. To try and combat this, mankind has, for a long time, tried to create repellents to keep the insects away. These repellents affect receptors in the mosquitos, keeping them at bay. One receptor that is highly expressed in the mosquito’s prey/host sensory organ is the Painless protein. By better understanding this receptor, more effective repellents might be available. This project delves into the structural studies of the protein by expressing it in Pichia pastoris and purifying it using a His tag and an attached GFP tag. The aim is to use Cryo-EM to determine its structure. Electrophysiology was also performed, showing that the protein does not react to some common TRP protein activators, yet is activated by H2O2. (Less)
Please use this url to cite or link to this publication:
author
Farkas, Titus LU
supervisor
organization
course
KEMR30 20242
year
type
H2 - Master's Degree (Two Years)
subject
keywords
Painless, Aedes aegypti, Structural Biology, Electrophysiology, Biochemistry
language
English
id
9182076
date added to LUP
2025-01-23 14:34:42
date last changed
2025-01-23 14:34:42
@misc{9182076,
  abstract     = {{Aedes aegypti is one of the most lethal disease vectors in modern history, spreading a majority of the deadliest diseases mediated by mosquitos. To try and combat this, mankind has, for a long time, tried to create repellents to keep the insects away. These repellents affect receptors in the mosquitos, keeping them at bay. One receptor that is highly expressed in the mosquito’s prey/host sensory organ is the Painless protein. By better understanding this receptor, more effective repellents might be available. This project delves into the structural studies of the protein by expressing it in Pichia pastoris and purifying it using a His tag and an attached GFP tag. The aim is to use Cryo-EM to determine its structure. Electrophysiology was also performed, showing that the protein does not react to some common TRP protein activators, yet is activated by H2O2.}},
  author       = {{Farkas, Titus}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{Expression of Aedes aegypti painless protein for structural studies : Production, solubilization, and purification of membrane proteins}},
  year         = {{2024}},
}