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@article{Kooistra:2018:10.1371/journal.pone.0209699,
author = {Kooistra, RL and David, R and Ruiz, AC and Powers, SW and Haselton, KJ and Kiernan, K and Blagborough, AM and Solamen, L and Olsen, KW and Putonti, C and Kanzok, SM},
doi = {10.1371/journal.pone.0209699},
journal = {PLoS One},
pages = {1--21},
title = {Characterization of a protozoan Phosducin-like protein-3 (PhLP-3) reveals conserved redox activity},
url = {http://dx.doi.org/10.1371/journal.pone.0209699},
volume = {13},
year = {2018}
}

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TY - JOUR
AB - We recently identified three novel thioredoxin-like genes in the genome of the protozoan parasite Plasmodium that belong to the Phosducin-like family of proteins (PhLP). PhLPs are small cytosolic proteins hypothesized to function in G-protein signaling and protein folding. Although PhLPs are highly conserved in eukaryotes from yeast to mammals, only a few representatives have been experimentally characterized to date. In addition, while PhLPs contain a thioredoxin domain, they lack a CXXC motif, a strong indicator for redox activity, and it is unclear whether members of the PhLP family are enzymatically active. Here, we describe PbPhLP-3 as the first phosducin-like protein of a protozoan organism, Plasmodium berghei. Initial transcription analysis revealed continuous low-level expression of pbphlp-3 throughout the complex Plasmodium life cycle. Attempts to knockout pbphlp-3 in P. berghei did not yield live parasites, suggesting an essential role for the gene in Plasmodium. We cloned, expressed and purified PbPhLP-3 and determined that the recombinant protein is redox active in vitro in a thioredoxin-coupled redox assay. It also has the capacity to reduce the organic compound tert-Butyl hydroperoxide (TBHP) in vitro, albeit at low efficiency. Sequence analysis, structural modeling, and site-directed mutagenesis revealed a conserved cysteine in the thioredoxin domain to be the redox active residue. Lastly, we provide evidence that recombinant human PhLP-3 exhibits redox activity similar to that of PbPhLP-3 and suggest that redox activity may be conserved in PhLP-3 homologs of other species. Our data provide new insight into the function of PhLP-3, which is hypothesized to act as co-chaperones in the folding and regulation of cytoskeletal proteins. We discuss the potential implications of PhLP-3 as a thioredoxin-target protein and possible links between the cellular redox network and the eukaryotic protein folding machinery.
AU - Kooistra,RL
AU - David,R
AU - Ruiz,AC
AU - Powers,SW
AU - Haselton,KJ
AU - Kiernan,K
AU - Blagborough,AM
AU - Solamen,L
AU - Olsen,KW
AU - Putonti,C
AU - Kanzok,SM
DO - 10.1371/journal.pone.0209699
EP - 21
PY - 2018///
SN - 1932-6203
SP - 1
TI - Characterization of a protozoan Phosducin-like protein-3 (PhLP-3) reveals conserved redox activity
T2 - PLoS One
UR - http://dx.doi.org/10.1371/journal.pone.0209699
UR - http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000454627200073&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
UR - https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0209699
UR - http://hdl.handle.net/10044/1/76725
VL - 13
ER -

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