Heterologous expression of proteins from Plasmodium falciparum: results from 1000 genes

Molecular and Biochemical Parasitology
Christopher MehlinWim G J Hol

Abstract

As part of a structural genomics initiative, 1000 open reading frames from Plasmodium falciparum, the causative agent of the most deadly form of malaria, were tested in an E. coli protein expression system. Three hundred and thirty-seven of these targets were observed to express, although typically the protein was insoluble. Sixty-three of the targets provided soluble protein in yields ranging from 0.9 to 406.6 mg from one liter of rich media. Higher molecular weight, greater protein disorder (segmental analysis, SEG), more basic isoelectric point (pI), and a lack of homology to E. coli proteins were all highly and independently correlated with difficulties in expression. Surprisingly, codon usage and the percentage of adenosines and thymidines (%AT) did not appear to play a significant role. Of those proteins which expressed, high pI and a hypothetical annotation were both strongly and independently correlated with insolubility. The overwhelmingly important role of pI in both expression and solubility appears to be a surprising and fundamental issue in the heterologous expression of P. falciparum proteins in E. coli. Twelve targets which did not express in E. coli from the native gene sequence were codon-optimized through whol...Continue Reading

Citations

Aug 25, 2009·Journal of Structural and Functional Genomics·Erin QuartleyElizabeth J Grayhack
Jan 12, 2011·Molecular Biotechnology·Charles GauciMarshall W Lightowlers
Oct 10, 2008·Nature·Elizabeth Ann Winzeler
Apr 7, 2010·Proceedings of the National Academy of Sciences of the United States of America·Itamar YadidDan S Tawfik
Sep 6, 2012·Acta Crystallographica. Section D, Biological Crystallography·Eric T LarsonEthan A Merritt
Mar 3, 2006·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Margaret A HolmesEthan A Merritt
May 10, 2006·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Margaret A HolmesEthan A Merritt
Mar 3, 2007·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Tracy ArakakiEthan A Merritt
Sep 10, 2011·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Jan AbendrothLance J Stewart
Sep 10, 2011·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Darren W BegleyLance J Stewart
Sep 10, 2011·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·David J LeiblyWesley C Van Voorhis
Sep 10, 2011·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Ryan ChoiWesley Van Voorhis
Sep 10, 2011·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·David J LeiblyWesley Van Voorhis
Sep 10, 2011·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Stephen N HewittWesley C Van Voorhis
Sep 10, 2011·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Douglas R DaviesLance J Stewart
Jul 23, 2008·Antimicrobial Agents and Chemotherapy·Kayode K OjoWesley C Van Voorhis
Feb 2, 2011·Antimicrobial Agents and Chemotherapy·Sayaka ShibataFrederick S Buckner
May 28, 2009·Journal of Biomolecular Screening·Gregory J CrowtherWesley C van Voorhis
May 24, 2007·Malaria Journal·Marni WilliamsLyn-Marie Birkholtz
Jan 1, 2011·Microbial Informatics and Experimentation·W Nicholson PriceJohn F Hunt
Sep 3, 2010·PLoS Neglected Tropical Diseases·Gregory J CrowtherFernán Agüero
Nov 11, 2011·PloS One·Erin J EngelsonWesley C Van Voorhis
Dec 14, 2012·Future Medicinal Chemistry·Isabelle S LucetChristian Doerig
Oct 17, 2013·Biotechnology Letters·Quinn Lu, Juan C Aon
Sep 10, 2014·Protein Expression and Purification·Inayat Hussain SheikhNuzhat A Kaushal
Oct 15, 2013·Eukaryotic Cell·Devaraja G Mudeppa, Pradipsinh K Rathod
Jan 24, 2016·Protein Expression and Purification·Laure BatailleElisabeth Garanger
Nov 1, 2009·Expert Opinion on Drug Discovery·Satoru TakeoTakafumi Tsuboi
Dec 23, 2015·Protein Expression and Purification·Mohd Shameel IqbalUday Bandyopadhyay
Feb 24, 2016·FEBS Letters·Sanna R RijpmaJan B Koenderink
Oct 16, 2015·Vaccine·Simon J DraperSumi Biswas
Oct 3, 2015·Vaccine·D Huw DaviesDenise L Doolan

❮ Previous
Next ❯

Related Concepts

Related Feeds

Antimalarial Agents (ASM)

Antimalarial agents, also known as antimalarials, are designed to prevent or cure malaria. Discover the latest research on antimalarial agents here.

Antimalarial Agents

Antimalarial agents, also known as antimalarials, are designed to prevent or cure malaria. Discover the latest research on antimalarial agents here.