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

Mehlin, Christopher; Boni, Erica; Buckner, Frederick S.; Engel, Linnea; Feist, Tiffany; Gelb, Michael H.; Haji, Lutfiyah; Kim, David; Liu, Colleen; Mueller, Natascha; Myler, Peter J.; Reddy, Joggari T.; Sampson, Joshua N.; Subramanian, E.; Voorhis, Wesley C. Van; Worthey, Elizabeth A.; Zucker, Frank; Hól, Wim G. J.

doi:10.1016/j.molbiopara.2006.03.011

Cited by 173 publications

(166 citation statements)

References 59 publications

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“…The above hypothesis is supported by analysis of the mutations that accumulated in the evolved variants Lib1-B7-18 and Lib2-D2-15. One factor to consider is the linkage between the protein's isoelectric point (pI) and soluble expression in E. coli (24). The evolved variant Lib1-B7-18 has a pI of 6.1, versus 8.9 for its starting point Lib1-B7.…”

Section: Resultsmentioning

confidence: 99%

Metamorphic proteins mediate evolutionary transitions of structure

Yadid

Kirshenbaum²,

Sharon³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

100

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The primary sequence of proteins usually dictates a single tertiary and quaternary structure. However, certain proteins undergo reversible backbone rearrangements. Such metamorphic proteins provide a means of facilitating the evolution of new folds and architectures. However, because natural folds emerged at the early stages of evolution, the potential role of metamorphic intermediates in mediating evolutionary transitions of structure remains largely unexplored. We evolved a set of new proteins based on ∼100 amino acid fragments derived from tachylectin-2-a monomeric, 236 amino acids, five-bladed β-propeller. Their structures reveal a unique pentameric assembly and novel β-propeller structures. Although identical in sequence, the oligomeric subunits adopt two, or even three, different structures that together enable the pentameric assembly of two propellers connected via a small linker. Most of the subunits adopt a wild-type-like structure within individual five-bladed propellers. However, the bridging subunits exhibit domain swaps and asymmetric strand exchanges that allow them to complete the two propellers and connect them. Thus, the modular and metamorphic nature of these subunits enabled dramatic changes in tertiary and quaternary structure, while maintaining the lectin function. These oligomers therefore comprise putative intermediates via which β-propellers can evolve from smaller elements. Our data also suggest that the ability of one sequence to equilibrate between different structures can be evolutionary optimized, thus facilitating the emergence of new structures.beta-propellers | conformational diversity | lectins | oligomerization | protein evolution M aynard-Smith's conjecture of protein evolution dictates that transitions of function and structure occur gradually (by single mutational steps), and smoothly, namely via intermediates that are all functional (1). This restriction imposes a major challenge, because mutational steps may create large structural perturbations that need to be tolerated while retaining function (2). In single domain proteins, significant "cut and paste" structural changes perturb a well-packed hydrophobic core (3, 4). Evolutionary transitions from one folded and functional domain structure into a new one should therefore (or must, by default, if one follows Maynard-Smith's conjecture) involve intermediates able to adopt more than one structure (5, 6). It has been also speculated that evolutionary intermediates of many existing folds, and folds that show internal symmetry in particular, were oligomeric assemblies of smaller subunits. Interestingly, most of the reported "metamorphic" proteins-proteins adopting more than one fold (7), are oliogomers in at least one of the two folds they adopt (8-10). Oligomerization may therefore serve as a means of augmenting structural metamorphism and of facilitating the emergence of new folds (11). However, metamorphic proteins are very rarely identified, and their evolutionary relevance is yet to be established. The involvement of struct...

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Section: Resultsmentioning

confidence: 99%

Metamorphic proteins mediate evolutionary transitions of structure

Yadid

Kirshenbaum²,

Sharon³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

100

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“…As part of a structural genomics initiative, 1000 open reading frames from P. falciparum were tested in an E. coli protein expression system. Three hundred and thirty-seven of the targets were expressed in E. coli [9]. For every soluble protein obtained, more than four were observed to express insolubly and more than 10 did not express at all.…”

Section: Discussionmentioning

confidence: 99%

“…Similar approaches have been used in high-throughput expression of various P. falciparum genes [9]. Since our intention was to compare the functional properties and cyclosporin-binding capacity of all of the cyclophilin/cyclophilin-like family members, we wished to obtain recombinant proteins under conditions as close to identical as possible.…”

mentioning

confidence: 99%

Overexpression, purification and assessment of cyclosporin binding of a family of cyclophilins and cyclophilin-like proteins of the human malarial parasite Plasmodium falciparum

Marín-Menéndez

Bell

2011

Protein Expression and Purification

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“…10 The protein was dissolved in ammonium acetate (10 mM, pH 6.8) to generate stock solution (4.5 µM).…”

Section: Enzyme Assaymentioning

confidence: 99%

Bioaffinity Mass Spectrometry Screening

Yang

Feng

et al. 2016

SLAS Discovery

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Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR-MS or ESI-FTMS) was used to screen 192 natural product extracts and a 659-member natural product-based fragment library for bindings to a potential malaria drug target, Plasmodium falciparum Rab11a (PfRab11a, PF13_0119). One natural product extract and 11 fragments showed binding activity. A new natural product, arborside E, was identified from the active extract of Psydrax montigena as a weak binder. Its binding activity and inhibitory activity against PfRab11a were confirmed by ESI-FTMS titration experiments and an orthogonal enzyme assay.

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

Cited by 173 publications

References 59 publications

Metamorphic proteins mediate evolutionary transitions of structure

Metamorphic proteins mediate evolutionary transitions of structure

Overexpression, purification and assessment of cyclosporin binding of a family of cyclophilins and cyclophilin-like proteins of the human malarial parasite Plasmodium falciparum

Bioaffinity Mass Spectrometry Screening

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