Mosquito Proteomics: Present and Future Prospective

Proteomics isthe detailed study of protein structure, function and protein-proteininteraction. In the few last years a lot of research work has been done in  genomes and transcriptomes by using highthroughput methods of analysis to understand the cellular functions. But nowthere is limited scope in the field of genomics and hence there is rapid moveto the proteomics. Proteomics is an integral part of system-level analysis ofcellular functions, promising to shed light on regulatory mechanisms that are beyondthe reach of genomics and transcriptomics. Proteomics is a rapidly evolving field due to advancements in thetechniques of identification of proteins by MS-spectrometry,  improvements in methods of protein-proteininteraction analysis, development of high-throughput approaches to synthesizeand purify proteins for microarray and crystallography applications. Currentproteomics approaches not only identify proteins expression, but alsoprotein-protein interactions and post-translational modifications. In thepresent review an attempt has been made to study proteomics analysis inmosquitoes.Key words: Anopheles, Aedes, Culex, Protein expression etc

<span style="mso-bidi-language:HI">Identification and characterization of a new putative c-type lysozyme <span style="mso-bidi-language:HI">from malaria vector <i>Anopheles stephensi</i> </span></span>

15-19Lysozyme (E.C. 3.<span style="color:black; mso-bidi-language:HI">2.1<span style="color:#242424;mso-bidi-language: HI">.17) activity is reported from the malaria vector Anopheles stephensi. The activity was detected in the salivary gland and midgut using bacteriolytic radial diffusion assay. Spectrophotometric analysis indicated that higher level of lysozyme activity was maintained in both midgut and salivary gland tissues. The activity reached the highest level in 4-8 days old mosquitoes. Genomic PCR amplification revealed the presence of at least two putative lysozyme genes in the mosquito genome. Preliminary analysis of one of the 413 bp genomic fragments showed 56% identity to the lysozyme of mosquito A. gambiae. However, the nature and origin of the putative cloned lysozyme gene remains elusive. </span

Effect of anti-fat body antibodies on reproductive capacity of mosquito <i style="">Anopheles stephensi</i> and transmission blocking of <i style="">Plasmodium vivax</i>

479-482Effect of anti-mosquito-fat body antibodies on the development of the malaria parasite, Plasmodium vivax has been studied by feeding Anopheles stephensi mosquitoes with infected blood supplemented with serum from immunized rabbits. Immunogenic polypeptides were identified by western blot. Mosquitoes that ingested anti-fat body antibodies along with infectious blood meal had significantly fewer oocysts than the mosquitoes in the control group. Effect of anti-mosquito fat body antibodies on fecundity, hatchability, mortality and engorgement of mosquitoes has also been reported. A significant reduction in fecundity and hatchability was observed, however, effect on mortality and engorgement was variable and statistically insignificant. Results indicated that fat body antibodies have the potential to disrupt reproductive physiology of malaria vector An. stephensi

Structural analysis of the <i>Pf</i>WDRs.

<p>a) Predicted structures of the 8 <i>Pf</i>WDRs by homology modeling with >90% confidence level and ≥95% residues in the allowed region of Ramachandran plot b) Predicted 3D structure of <i>Pf</i>CAF-1 subunit (PF3D7_0110700) depicting histone H4 binding residues (yellow) as inferred from its human homolog RBBP4/RBBP7. c) Structure of <i>Hs</i>RBBP7 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref005" target="_blank">5</a>] [PDB: 3CFV, green] highlighting H4 binding residues (magenta). d) Superimposition of structures of <i>Pf</i>CAF-1 subunit and <i>Hs</i>RBBP7 with histone H4 peptide (red) clearly showing overlapping histone H4 binding pockets (highlighted in dotted circle). Close-up view of overlapped histone binding pockets is also shown depicting variant residues (highlighted as sticks) of <i>Pf</i>CAF-1 in comparison to <i>Hs</i>RBBP7. Residues position in the figure are according to <i>Hs</i>RBBP7 e.g. F29L represents Phe at 29^th position of <i>Hs</i>RBBP7 is replaced by Leu in <i>Pf</i>CAF-1. e) Overlay of <i>Pf</i>CAF-1 model (light blue) and <i>Hs</i>RBBP4 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref006" target="_blank">6</a>] crystal structure [PDB: 2XU7, green] highlighting FOG-1 binding residues as yellow and red sticks respectively. Residues position scheme as mentioned above. f) Structural alignment of 3D model of <i>Pf</i>RACK (PF3D7_0826700-light blue) and <i>Hs</i>RACK1 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref007" target="_blank">7</a>] [PDB: 4AOW, green]. The residues of hydrophobic ring important in binding to protein ligands at the top surface of propeller structure are shown as yellow and red sticks for <i>Pf</i>RACK and <i>Hs</i>RACK1 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref007" target="_blank">7</a>], respectively. Insertions in <i>Pf</i>RACK are highlighted in red that mainly lie in the loop regions. g) Overlay of predicted model of <i>Pf</i>WDR92 (PF3D7_1347000-light blue) with the crystal structure of <i>Hs</i>WDR92/Monad [PDB: 3I2N, green] comparing loops with insertion in <i>P</i>. <i>falciparum</i> i.e. <i>Pf</i> long loop (red) and <i>Hs</i> short loop (cyan). h) A structure based sequence alignment between <i>Pf</i>CAF-1 and <i>Hs</i>RBBP7. Secondary structure elements of <i>Hs</i>RBBP7 are shown below the alignment indicated by coils, arrows and gaps for helices, β-strands and loops, respectively [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref005" target="_blank">5</a>]. Green star and magenta boxes above the alignment indicate key residues involved in hydrophobic and hydrophilic interactions with histone H4, respectively [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref005" target="_blank">5</a>]. Conserved residues are highlighted in yellow boxes while similar residues are highlighted in green text. Black dotted line below alignment indicates sequence part for which no structure is available.</p

Expression patterns of the <i>Pf</i>WDR genes during life cycle of the parasite.

<p>A <i>Pf</i>WDRs phaseogram from microarray data of Llinas/Derisi et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref025" target="_blank">25</a>] was generated covering IDC (1–48h) and compared with Le Roch/Winzeler et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref026" target="_blank">26</a>] data from two independently synchronized <i>P</i>. <i>falciparum</i> 3D7 cultures i.e temperature and sorbitol covering IDC stages (R,T,S,M) as well as G and Sp. Colorimetric representation used for heat maps of transcriptome data is green-red (green, low expression; black, medium expression; red, high expression). Heat map panels at the right side with blue-red colour scale (blue, low expression; red, high expression) represent comparison of proteome and phosphoproteome data obtained from (<b>a</b>) Florens et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref027" target="_blank">27</a>], (<b>b</b> & <b>c</b>) Lasonder et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref028" target="_blank">28</a>,<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref029" target="_blank">29</a>], (<b>d</b>) Le Roch et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref030" target="_blank">30</a>], (<b>e</b>) Khan et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref031" target="_blank">31</a>], (<b>f</b>) Silvestrini et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref032" target="_blank">32</a>], (<b>g</b>) Oehring et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref033" target="_blank">33</a>], (<b>h</b>) Linder et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref034" target="_blank">34</a>], (<b>i</b>) Solyakov et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref035" target="_blank">35</a>], (<b>j</b>) Treeck et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref036" target="_blank">36</a>] and (<b>k</b>-I & <b>k</b>-II) Pease et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref037" target="_blank">37</a>]. Column to the right indicates PlasmoDB gene IDs of <i>Pf</i>WDRs coloured according to the functional classification (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.g005" target="_blank">Fig 5</a>). Different life cycle stages are abbreviated as: ER and LR, early and late rings; ET and LT, early and late trophozoites; ES and LS, early and late schizonts; M, merozoites; G, gametocytes; Sp, sporozoites; Gt, gamete; EG, early gametocyte; MG, mature gametocyte; OOC, oocyst; ODS, oocyst derived sporozoites; SGS, salivary gland sporozoites; phosEnr, phospho-enriched; phosDep, phospho-depleted; Nuc, nuclear; and cyto, cytoplasmic. Grey colour represents absence of detection.</p

Sequence logos for the WD40 motif.

<p>a) The <i>Pf</i>WDRs HMM logo based on alignment of all the identified WDRs in <i>P</i>. <i>falciparum</i>. b) The Pfam WD40 family HMM logo drawn from alignment of Pfam WD40 seed sequences.</p

Physical mapping of <i>Pf</i>WDR genes depicting their genomic localization onto 14 chromosomes of <i>P</i>. <i>falciparum</i>.

<p>Positions of centromeres are represented by filled circles on the chromosomes (vertical bars). Integers at the top of each bar indicate chromosome number. Grouped genes, adjacent genes and genes leaving one or two gene positions in between; are highlighted with black and red asterisks, respectively. Further, asterisks for clusters having co-expressed genes are encircled. Genes on chromosomes are colour coded as per their functional classification (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.g005" target="_blank">Fig 5</a>). The scale on the left is in megabases (Mb). Number of <i>Pf</i>WDR genes per chromosome is also shown in the graph.</p

Genome-Wide Collation of the <i>Plasmodium falciparum</i> WDR Protein Superfamily Reveals Malarial Parasite-Specific Features

<div><p>Despite a significant drop in malaria deaths during the past decade, malaria continues to be one of the biggest health problems around the globe. WD40 repeats (WDRs) containing proteins comprise one of the largest and functionally diverse protein superfamily in eukaryotes, acting as scaffolds for assembling large protein complexes. In the present study, we report an extensive <i>in silico</i> analysis of the WDR gene family in human malaria parasite <i>Plasmodium falciparum</i>. Our genome-wide identification has revealed 80 putative WDR genes in <i>P</i>. <i>falciparum</i> (<i>Pf</i>WDRs). Five distinct domain compositions were discovered in <i>Plasmodium</i> as compared to the human host. Notably, 31 <i>Pf</i>WDRs were annotated/re-annotated on the basis of their orthologs in other species. Interestingly, most <i>Pf</i>WDRs were larger as compared to their human homologs highlighting the presence of parasite-specific insertions. Fifteen <i>Pf</i>WDRs appeared specific to the <i>Plasmodium</i> with no assigned orthologs. Expression profiling of <i>Pf</i>WDRs revealed a mixture of linear and nonlinear relationships between transcriptome and proteome, and only nine <i>Pf</i>WDRs were found to be stage-specific. Homology modeling identified conservation of major binding sites in <i>Pf</i>CAF-1 and <i>Pf</i>RACK. Protein-protein interaction network analyses suggested that <i>Pf</i>WDRs are highly connected proteins with ~1928 potential interactions, supporting their role as hubs in cellular networks. The present study highlights the roles and relevance of the WDR family in <i>P</i>. <i>falciparum</i>, and identifies unique features that lay a foundation for further experimental dissection of <i>Pf</i>WDRs.</p></div

Extraction and characterization of the <i>Pf</i>WDRs.

<p>a) Schematic representation of the approaches employed for the identification of <i>Pf</i>WDR genes. b) Graphical representation of the occurrence of introns by number in <i>Pf</i>WDR genes. c) Predicted percentage of the proteome of eukaryotic organisms devoted for the WDR proteins. Apicomplexans are boxed. d) Distribution of the WD40 motifs by number in <i>Pf</i>WDRs.</p

Protein-protein interactions (PPIs) network analysis for the <i>Pf</i>WDRs.

<p>a) PPIs network of all the 80 <i>Pf</i>WDRs (yellow nodes). b) PPIs network of the <i>Pf</i>WDRs predicted to be engaged in chromatin assembly and remodeling (yellow nodes). Node size is proportional to the degree of node. Nodes are coloured according to their functional classification based on PlasmoDB/human homologs annotations. Edge width is proportional to the confidence score from STRING for each interaction. Interactions among <i>Pf</i>WDR proteins are highlighted with red edges. Nodes not coexpressed even at a single stage with the <i>Pf</i>WDRs are encircled in red. The nodes for which no protein expression data was available at PlasmoDB are encircled in blue colour. c) PPIs network of <i>Pf</i>Sec13 (PF3D7_1230700) (yellow and magenta node) derived from STRING (outer ring with blue edges), co-IP [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128507#pone.0128507.ref016" target="_blank">16</a>] (inner ring with orange edges) and Y2H (triangles with green edges). Interactions common between STRING and co-IP are indicated by diamond shapes and black edges. Nodes are colour coded as per their functions.</p