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The Biological Bulletin Jun 2016Barnacles permanently adhere to nearly any inert substrate using proteinaceous glue. The glue consists of at least ten major proteins, some of which have been isolated...
Barnacles permanently adhere to nearly any inert substrate using proteinaceous glue. The glue consists of at least ten major proteins, some of which have been isolated and sequenced. Questions still remain about the chemical mechanisms involved in adhesion and the potential of the glue to serve as a platform for mineralization of the calcified base plate. We tested the hypothesis that barnacle glue contains phosphoproteins, which have the potential to play a role in both adhesion and mineralization. Using a combination of phosphoprotein-specific gel staining and Western blotting with anti-phosphoserine antibody, we identified multiple phosphorylated proteins in uncured glue secretions from the barnacle Amphibalanus amphitrite The protein composition of the glue and the quantity and abundance of phosphoproteins varied distinctly among individual barnacles, possibly due to cyclical changes in the glue secretion over time. We assessed the location of the phosphoproteins within the barnacle glue layer using decalcified barnacle base plates and residual glue deposited by reattached barnacles. Phosphoproteins were found throughout the organic matrix of the base plate and within the residual glue. Staining within the residual glue appeared most intensely in regions where capillary glue ducts, which are involved in cyclical release of glue, had been laid down. Lastly, mineralization studies of glue proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) indicated that proteins identified as phosphorylated possibly induce mineralization of calcium carbonate (CaCO3). These results contribute to our understanding of the protein composition of barnacle glue, and provide new insights into the potential roles of phosphoproteins in underwater bioadhesives.
Topics: Adhesives; Animals; Blotting, Western; Calcium Carbonate; Phosphoproteins; Thoracica
PubMed: 27365418
DOI: 10.1086/BBLv230n3p233 -
Journal of Virology Nov 2021Respiratory syncytial virus (RSV) is a single-stranded, negative-sense RNA virus in the family and genus that can cause severe disease in infants, immunocompromised...
Respiratory syncytial virus (RSV) is a single-stranded, negative-sense RNA virus in the family and genus that can cause severe disease in infants, immunocompromised adults, and the elderly. The RSV viral RNA-dependent RNA polymerase (vRdRp) complex is composed of the phosphoprotein (P) and the large polymerase protein (L). The P protein is constitutively phosphorylated by host kinases and has 41 serine (S) and threonine (T) residues as potential phosphorylation sites. To identify important phosphorylation residues in the P protein, we systematically and individually mutated all S and T residues to alanine (A) and analyzed their effects on genome transcription and replication by using a minigenome system. We found that the mutation of eight residues resulted in minigenome activity significantly lower than that of wild-type (WT) P. We then incorporated these mutations (T210A, S203A, T151A, S156A, T160A, S23A, T188A, and T105A) into full-length genome cDNA to rescue recombinant RSV. We were able to recover four recombinant viruses (with T151A, S156A, T160A, or S23A), suggesting that RSV-P residues T210, S203, T188, and T105 are essential for viral RNA replication. Among the four recombinant viruses rescued, rRSV-T160A caused a minor growth defect relative to its parental virus while rRSV-S156A had severely restricted replication due to decreased levels of genomic RNA. During infection, P-S156A phosphorylation was decreased, and when passaged, the S156A virus acquired a known compensatory mutation in L (L795I) that enhanced both WT-P and P-S156A minigenome activity and was able to partially rescue the S156A viral growth defect. This work demonstrates that residues T210, S203, T188, and T105 are critical for RSV replication and that S156 plays a critical role in viral RNA synthesis. RSV-P is a heavily phosphorylated protein that is required for RSV replication. In this study, we identified several residues, including P-S156, as phosphorylation sites that play critical roles in efficient viral growth and genome replication. Future studies to identify the specific kinase(s) that phosphorylates these residues can lead to kinase inhibitors and antiviral drugs for this important human pathogen.
Topics: Animals; Chlorocebus aethiops; Genome, Viral; Phosphoproteins; RNA, Viral; Respiratory Syncytial Virus, Human; Transcription, Genetic; Vero Cells; Viral Proteins; Virus Replication
PubMed: 34613802
DOI: 10.1128/JVI.01206-21 -
Life Sciences Oct 2021Betacoronaviruses are in one genera of coronaviruses including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), severe acute respiratory syndrome... (Review)
Review
Betacoronaviruses are in one genera of coronaviruses including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome-related coronavirus (MERS-CoV), etc. These viruses threaten public health and cause dramatic economic losses. The nucleocapsid (N) protein is a structural protein of betacoronaviruses with multiple functions such as forming viral capsids with viral RNA, interacting with viral membrane protein to form the virus core with RNA, binding to several cellular kinases for signal transductions, etc. In this review, we highlighted the potential of the N protein as a suitable antiviral target from different perspectives, including structure, functions, and antiviral strategies for combatting betacoronaviruses.
Topics: Animals; Antiviral Agents; Betacoronavirus; Coronavirus Infections; Coronavirus Nucleocapsid Proteins; Drug Discovery; Host-Pathogen Interactions; Humans; Middle East Respiratory Syndrome Coronavirus; Models, Molecular; Molecular Targeted Therapy; Phosphoproteins; Protein Interaction Maps; SARS-CoV-2; COVID-19 Drug Treatment
PubMed: 33189817
DOI: 10.1016/j.lfs.2020.118754 -
RNA Biology 2010The phosphoprotein P of non-segmented negative-sense RNA viruses is an essential component of the replication and transcription complex and acts as a co-factor for the... (Review)
Review
The structure of the nucleoprotein binding domain of lyssavirus phosphoprotein reveals a structural relationship between the N-RNA binding domains of Rhabdoviridae and Paramyxoviridae.
The phosphoprotein P of non-segmented negative-sense RNA viruses is an essential component of the replication and transcription complex and acts as a co-factor for the viral RNA-dependent RNA polymerase. P recruits the viral polymerase to the nucleoprotein-bound viral RNA (N-RNA) via an interaction between its C-terminal domain and the N-RNA complex. We have obtained the structure of the C-terminal domain of P of Mokola virus (MOKV), a lyssavirus that belongs to the Rhabdoviridae family and mapped at the amino acid level the crucial positions involved in interaction with N and in the formation of the viral replication complex. Comparison of the N-RNA binding domains of P solved to date suggests that the N-RNA binding domains are structurally conserved among paramyxoviruses and rhabdoviruses in spite of low sequence conservation. We also review the numerous other functions of this domain and more generally of the phosphoprotein.
Topics: Humans; Lyssavirus; Models, Biological; Models, Molecular; Nucleoproteins; Paramyxoviridae; Phosphoproteins; Protein Structure, Tertiary; RNA; RNA-Binding Proteins; Rhabdoviridae; Structure-Activity Relationship
PubMed: 20458178
DOI: 10.4161/rna.7.3.11931 -
Biomolecular NMR Assignments Oct 2020Protein Phosphatase 2A, PP2A, the principal Serine/threonine phosphatase, has major roles in broad range of signaling pathways that include regulation of cell cycle,...
Protein Phosphatase 2A, PP2A, the principal Serine/threonine phosphatase, has major roles in broad range of signaling pathways that include regulation of cell cycle, cell proliferation and neuronal signaling. The loss of function of PP2A is linked with many human diseases, like cancer and neurodegenerative disorders. Protein phosphatase 2A (PP2A) functions as tumor suppressor and its tumor suppressor activity is inhibited by the overexpression of PP2A inhibitor proteins in most of the cancers. ARPP-19/ARPP-16 has been identified as one of the potential PP2A inhibitor proteins. Here, we report the resonance assignment of backbone H, C and N atoms of human ARPP-19 and ARPP-16 proteins. These chemical shift values can provide valuable information for the further study of the dynamics and interaction of ARPP-proteins to PP2A using NMR spectroscopy.
Topics: Amino Acid Sequence; Carbon-13 Magnetic Resonance Spectroscopy; Humans; Nitrogen Isotopes; Phosphoproteins; Proton Magnetic Resonance Spectroscopy
PubMed: 32468417
DOI: 10.1007/s12104-020-09951-w -
International Journal of Molecular... Oct 2017Drought is one of the major abiotic stresses that negatively affects plant growth and development. is an ecologically important shrub in the mid-Asia desert region and...
Drought is one of the major abiotic stresses that negatively affects plant growth and development. is an ecologically important shrub in the mid-Asia desert region and used as a model for abiotic tolerance research in trees. Protein phosphorylation participates in the regulation of various biological processes, however, phosphorylation events associated with drought stress signaling and response in plants is still limited. Here, we conducted a quantitative phosphoproteomic analysis of the response of roots to short-term drought stress. Data are available via the iProx database with project ID IPX0000971000. In total, 7841 phosphorylation sites were found from the 2019 identified phosphopeptides, corresponding to 1060 phosphoproteins. Drought stress results in significant changes in the abundance of 103 phosphopeptides, corresponding to 90 differentially-phosphorylated phosphoproteins (DPPs). Motif-x analysis identified two motifs, including [pSP] and [RXXpS], from these DPPs. Functional enrichment and protein-protein interaction analysis showed that the DPPs were mainly involved in signal transduction and transcriptional regulation, osmotic adjustment, stress response and defense, RNA splicing and transport, protein synthesis, folding and degradation, and epigenetic regulation. These drought-corresponsive phosphoproteins, and the related signaling and metabolic pathways probably play important roles in drought stress signaling and response in roots. Our results provide new information for understanding the molecular mechanism of the abiotic stress response in plants at the posttranslational level.
Topics: Amino Acid Motifs; Amino Acid Sequence; Computational Biology; Droughts; Fabaceae; Phosphoproteins; Plant Proteins; Plant Roots; Protein Interaction Mapping; Protein Interaction Maps; Proteome; Proteomics; Stress, Physiological; Tandem Mass Spectrometry
PubMed: 29039783
DOI: 10.3390/ijms18102158 -
Biosensors Oct 2022Abnormal protein phosphorylation may relate to diseases such as Alzheimer's, schizophrenia, and Parkinson's. Therefore, the real-time detection of phosphoproteins in...
Abnormal protein phosphorylation may relate to diseases such as Alzheimer's, schizophrenia, and Parkinson's. Therefore, the real-time detection of phosphoproteins in sweat was of great significance for the early knowledge, detection, and treatment of neurological diseases. In this work, anatase/rutile TiO was in situ grown on the MXene surface to constructing the intercalation structure MXene@anatase/rutile TiO ternary heterostructure as a sensing platform for detecting phosphoprotein in sweat. Here, the intercalation structure of MXene acted as electron and diffusion channels for phosphoproteins. The in situ grown anatase/rutile TiO with n-n-type heterostructure provided specific adsorption sites for the phosphoproteins. The determination of phosphoprotein covered concentrations in sweat, with linear range from 0.01 to 1 mg/mL, along with a low LOD of 1.52 μM. It is worth noting that, since the macromolecular phosphoprotein was adsorbed on the surface of the material, the electrochemical signal gradually decreased with the increase of phosphoprotein concentration. In addition, the active sites in the MXene@anatase/rutile TiO ternary heterojunction and synergistic effect of the heterojunction were verified by first-principle calculations to further realize the response to phosphoproteins. Additionally, the effective diffusion capacity and mobility of phosphoprotein molecules in the ternary heterojunction structure were studied by molecular dynamics simulation. Furthermore, the constructed sensing platform showed high selectivity, repeatability, reproducibility, and stability, and this newly developed sensor can detect for phosphoprotein in actual sweat samples. This satisfactory sensing strategy could be promoted to realize the noninvasive and continuous detection of sweat.
Topics: Sweat; Phosphoproteins; Reproducibility of Results; Titanium
PubMed: 36291003
DOI: 10.3390/bios12100865 -
Scientific Reports Jan 2021Mutations of Odontogenesis-Associated Phosphoprotein (ODAPH, OMIM *614829) cause autosomal recessive amelogenesis imperfecta, however, the function of ODAPH during...
Mutations of Odontogenesis-Associated Phosphoprotein (ODAPH, OMIM *614829) cause autosomal recessive amelogenesis imperfecta, however, the function of ODAPH during amelogenesis is unknown. Here we characterized normal Odaph expression by in situ hybridization, generated Odaph truncation mice using CRISPR/Cas9 to replace the TGC codon encoding Cys41 into a TGA translation termination codon, and characterized and compared molar and incisor tooth formation in Odaph, Odaph, and Odaph mice. We also searched genomes to determine when Odaph first appeared phylogenetically. We determined that tooth development in Odaph and Odaph mice was indistinguishable in all respects, so the condition in mice is inherited in a recessive pattern, as it is in humans. Odaph is specifically expressed by ameloblasts starting with the onset of post-secretory transition and continues until mid-maturation. Based upon histological and ultrastructural analyses, we determined that the secretory stage of amelogenesis is not affected in Odaph mice. The enamel layer achieves a normal shape and contour, normal thickness, and normal rod decussation. The fundamental problem in Odaph mice starts during post-secretory transition, which fails to generate maturation stage ameloblasts. At the onset of what should be enamel maturation, a cyst forms that separates flattened ameloblasts from the enamel surface. The maturation stage fails completely.
Topics: Ameloblasts; Amelogenesis; Amelogenesis Imperfecta; Animals; Dental Enamel; Extracellular Matrix Proteins; Gene Knock-In Techniques; In Situ Hybridization; Incisor; Mice; Molar; Odontogenesis; Phosphoproteins
PubMed: 33441959
DOI: 10.1038/s41598-020-80912-y -
Scientific Reports Oct 2015Retinitis pigmentosa (RP) shows progressive loss of photoreceptors involved with heterogeneous genetic background. Here, by exome sequencing and linkage analysis on a...
Retinitis pigmentosa (RP) shows progressive loss of photoreceptors involved with heterogeneous genetic background. Here, by exome sequencing and linkage analysis on a Chinese family with autosomal dominant RP, we identified a putative pathogenic variant, p.Gly97Arg, in the gene SPP2, of which expression was detected in multiple tissues including retina. The p.Gly97Arg was absent in 800 ethnically matched chromosomes and 1400 in-house exome dataset, and was located in the first of the two highly conserved disulfide bonded loop of secreted phosphoprotein 2 (Spp-24) encoded by SPP2. Overexpression of p.Gly97Arg and another signal peptide mutation, p.Gly29Asp, caused cellular retention of both endogenous wild type and exogenous mutants in vitro, and primarily affected rod photoreceptors in zebrafish mimicking cardinal feature of RP. Taken together, our data indicate that the two mutations of SPP2 have dominant negative effects and cellular accumulation of Spp-24 might be particularly toxic to photoreceptors and/or retinal pigment epithelium. SPP2 has a new role in retinal degeneration.
Topics: Adult; Alleles; Animals; DNA Mutational Analysis; Electroretinography; Endoplasmic Reticulum; Gene Expression; Genes, Dominant; Genetic Association Studies; HEK293 Cells; Heterozygote; Humans; Male; Middle Aged; Mutation; Phenotype; Phosphoproteins; Photoreceptor Cells; Protein Transport; Retinitis Pigmentosa; Tomography, Optical Coherence; Young Adult; Zebrafish
PubMed: 26459573
DOI: 10.1038/srep14867 -
Developmental Cell Dec 2007A recent Neuron paper by Kwiatkowski et al. that analyzes the phenotypes of mouse embryos lacking all members of the Ena/VASP family reveals new roles for these... (Review)
Review
A recent Neuron paper by Kwiatkowski et al. that analyzes the phenotypes of mouse embryos lacking all members of the Ena/VASP family reveals new roles for these molecules in cortical development.
Topics: Animals; Cell Adhesion Molecules; Cytoskeletal Proteins; Malformations of Cortical Development; Mice; Mice, Knockout; Microfilament Proteins; Multigene Family; Phosphoproteins
PubMed: 18061556
DOI: 10.1016/j.devcel.2007.11.007