Matrix Protein Interactions with Synthetic Surfaces

2015 ◽  
pp. 91-146 ◽  
Author(s):  
Patricia Rico ◽  
Marco Cantini ◽  
George Altankov ◽  
Manuel Salmerón-Sánchez
Keyword(s):  
Protein Interactions ◽  
Matrix Protein

scholarly journals The Minor Matrix Protein VP24 from Ebola Virus Lacks Direct Lipid-Binding Properties

Viruses ◽  
2020 ◽  
Vol 12 (8) ◽  
pp. 869
Author(s):  
Yuan Su ◽  
Robert V. Stahelin
Keyword(s):  
Protein Interactions ◽  
Matrix Protein ◽  
Ebola Virus ◽  
Cellular Membrane ◽  
Lipid Binding ◽  
Confocal Imaging ◽  
Host Cells ◽  
Binding Ability ◽  
Matrix Layer

Viral protein 24 (VP24) from Ebola virus (EBOV) was first recognized as a minor matrix protein that associates with cellular membranes. However, more recent studies shed light on its roles in inhibiting viral genome transcription and replication, facilitating nucleocapsid assembly and transport, and interfering with immune responses in host cells through downregulation of interferon (IFN)-activated genes. Thus, whether VP24 is a peripheral protein with lipid-binding ability for matrix layer recruitment has not been explored. Here, we examined the lipid-binding ability of VP24 with a number of lipid-binding assays. The results indicated that VP24 lacked the ability to associate with lipids tested regardless of VP24 posttranslational modifications. We further demonstrate that the presence of the EBOV major matrix protein VP40 did not promote VP24 membrane association in vitro or in cells. Further, no protein–protein interactions between VP24 and VP40 were detected by co-immunoprecipitation. Confocal imaging and cellular membrane fractionation analyses in human cells suggested VP24 did not specifically localize at the plasma membrane inner leaflet. Overall, we provide evidence that EBOV VP24 is not a lipid-binding protein and its presence in the viral matrix layer is likely not dependent on direct lipid interactions.

HIV‐1 Matrix Protein Interactions with Cellular tRNAs

2020 ◽  
Vol 34 (S1) ◽  
pp. 1-1
Author(s):  
Jason Ejimogu ◽  
Connor Parker ◽  
Mahlet Bauerle ◽  
Cheyenne Palm ◽  
Janae Baptiste Brown ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Matrix Protein ◽  
Hiv 1

scholarly journals Complete Genome Sequence and In Planta Subcellular Localization of Maize Fine Streak Virus Proteins

2005 ◽  
Vol 79 (9) ◽  
pp. 5304-5314 ◽  
Author(s):  
Chi-Wei Tsai ◽  
Margaret G. Redinbaugh ◽  
Kristen J. Willie ◽  
Sharon Reed ◽  
Michael Goodin ◽  
...  
Keyword(s):  
Genome Sequence ◽  
Protein Interactions ◽  
Matrix Protein ◽  
Fluorescent Protein ◽  
Streak Virus ◽  
In Planta ◽  
Nuclear Localization Signals ◽  
N Protein ◽  
P Protein ◽  
Maize Fine Streak Virus

ABSTRACT The genome of the nucleorhabdovirus maize fine streak virus (MFSV) consists of 13,782 nucleotides of nonsegmented, negative-sense, single-stranded RNA. The antigenomic strand consisted of seven open reading frames (ORFs), and transcripts of all ORFs were detected in infected plants. ORF1, ORF6, and ORF7 had significant similarities to the nucleocapsid protein (N), glycoprotein (G), and polymerase (L) genes of other rhabdoviruses, respectively, whereas the ORF2, ORF3, ORF4, and ORF5 proteins had no significant similarities. The N (ORF1), ORF4, and ORF5 proteins localized to nuclei, consistent with the presence of nuclear localization signals (NLSs) in these proteins. ORF5 likely encodes the matrix protein (M), based on its size, the position of its NLS, and the localization of fluorescent protein fusions to the nucleus. ORF2 probably encodes the phosphoprotein (P) because, like the P protein of Sonchus yellow net virus (SYNV), it was spread throughout the cell when expressed alone but was relocalized to a subnuclear locus when coexpressed with the MFSV N protein. Unexpectedly, coexpression of the MFSV N and P proteins, but not the orthologous proteins of SYNV, resulted in accumulations of both proteins in the nucleolus. The N and P protein relocalization was specific to cognate proteins of each virus. The subcellular localizations of the MFSV ORF3 and ORF4 proteins were distinct from that of the SYNV sc4 protein, suggesting different functions. To our knowledge, this is the first comparative study of the cellular localizations of plant rhabdoviral proteins. This study indicated that plant rhabdoviruses are diverse in genome sequence and viral protein interactions.

scholarly journals Membrane Binding of HIV-1 Matrix Protein: Dependence on Bilayer Composition and Protein Lipidation

2016 ◽  
Vol 90 (9) ◽  
pp. 4544-4555 ◽  
Author(s):  
Marilia Barros ◽  
Frank Heinrich ◽  
Siddhartha A. K. Datta ◽  
Alan Rein ◽  
Ioannis Karageorgos ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Protein Interactions ◽  
Matrix Protein ◽  
Full Length ◽  
Protein Protein Interactions ◽  
Binding Studies ◽  
Protein Protein Interaction ◽  
Gag Polyprotein ◽  
Protein Lipidation ◽  
Hiv 1

ABSTRACTBy assembling in a protein lattice on the host's plasma membrane, the retroviral Gag polyprotein triggers formation of the viral protein/membrane shell. The MA domain of Gag employs multiple signals—electrostatic, hydrophobic, and lipid-specific—to bring the protein to the plasma membrane, thereby complementing protein-protein interactions, located in full-length Gag, in lattice formation. We report the interaction of myristoylated and unmyristoylated HIV-1 Gag MA domains with bilayers composed of purified lipid components to dissect these complex membrane signals and quantify their contributions to the overall interaction. Surface plasmon resonance on well-defined planar membrane models is used to quantify binding affinities and amounts of protein and yields free binding energy contributions, ΔG, of the various signals. Charge-charge interactions in the absence of the phosphatidylinositide PI(4,5)P2attract the protein to acidic membrane surfaces, and myristoylation increases the affinity by a factor of 10; thus, our data do not provide evidence for a PI(4,5)P2trigger of myristate exposure. Lipid-specific interactions with PI(4,5)P2, the major signal lipid in the inner plasma membrane, increase membrane attraction at a level similar to that of protein lipidation. While cholesterol does not directly engage in interactions, it augments protein affinity strongly by facilitating efficient myristate insertion and PI(4,5)P2binding. We thus observe that the isolated MA protein, in the absence of protein-protein interaction conferred by the full-length Gag, binds the membrane with submicromolar affinities.IMPORTANCELike other retroviral species, the Gag polyprotein of HIV-1 contains three major domains: the N-terminal, myristoylated MA domain that targets the protein to the plasma membrane of the host; a central capsid-forming domain; and the C-terminal, genome-binding nucleocapsid domain. These domains act in concert to condense Gag into a membrane-bounded protein lattice that recruits genomic RNA into the virus and forms the shell of a budding immature viral capsid. In binding studies of HIV-1 Gag MA to model membranes with well-controlled lipid composition, we dissect the multiple interactions of the MA domain with its target membrane. This results in a detailed understanding of the thermodynamic aspects that determine membrane association, preferential lipid recruitment to the viral shell, and those aspects of Gag assembly into the membrane-bound protein lattice that are determined by MA.

scholarly journals Analysis of Procollagen C-Proteinase Enhancer-1/Glycosaminoglycan Binding Sites and of the Potential Role of Calcium Ions in the Interaction

2019 ◽  
Vol 20 (20) ◽  
pp. 5021 ◽  
Author(s):  
Potthoff ◽  
Bojarski ◽  
Kohut ◽  
Lipska ◽  
Liwo ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Calcium Ions ◽  
Binding Sites ◽  
Matrix Protein ◽  
Structural Model ◽  
Extracellular Matrix Protein ◽  
Binding Assays ◽  
Gag Protein ◽  
Modeling Approaches ◽  
Computational Data

In this study, we characterize the interactions between the extracellular matrix protein, procollagen C-proteinase enhancer-1 (PCPE-1), and glycosaminoglycans (GAGs), which are linear anionic periodic polysaccharides. We applied molecular modeling approaches to build a structural model of full-length PCPE-1, which is not experimentally available, to predict GAG binding poses for various GAG lengths, types and sulfation patterns, and to determine the effect of calcium ions on the binding. The computational data are analyzed and discussed in the context of the experimental results previously obtained using surface plasmon resonance binding assays. We also provide experimental data on PCPE-1/GAG interactions obtained using inhibition assays with GAG oligosaccharides ranging from disaccharides to octadecasaccharides. Our results predict the localization of GAG-binding sites at the amino acid residue level onto PCPE-1 and is the first attempt to describe the effects of ions on protein-GAG binding using modeling approaches. In addition, this study allows us to get deeper insights into the in silico methodology challenges and limitations when applied to GAG-protein interactions.

scholarly journals Determination of protein regions responsible for interactions of amelogenin with CD63 and LAMP1

2007 ◽  
Vol 408 (3) ◽  
pp. 347-354 ◽  
Author(s):  
YanMing Zou ◽  
HongJun Wang ◽  
Jason L. Shapiro ◽  
Curtis T. Okamoto ◽  
Steven J. Brookes ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Matrix Protein ◽  
Cultured Cells ◽  
Matrix Proteins ◽  
Amino Acid Residues ◽  
Enamel Matrix ◽  
Protein Protein Interactions ◽  
Binding Region ◽  
Enamel Matrix Proteins

The enamel matrix protein amelogenin is secreted by ameloblasts into the extracellular space to guide the formation of highly ordered hydroxyapatite mineral crystallites, and, subsequently, is almost completely removed during mineral maturation. Amelogenin interacts with the transmembrane proteins CD63 and LAMP (lysosome-associated membrane protein) 1, which are involved in endocytosis. Exogenously added amelogenin has been observed to move rapidly into CD63/LAMP1-positive vesicles in cultured cells. In the present study, we demonstrate the protein region defined by amino acid residues 103–205 for CD63 interacts not only with amelogenin, but also with other enamel matrix proteins (ameloblastin and enamelin). A detailed characterization of binding regions in amelogenin, CD63 and LAMP1 reveals that the amelogenin region defined by residues PLSPILPELPLEAW is responsible for the interaction with CD63 through residues 165–205, with LAMP1 through residues 226–251, and with the related LAMP2 protein through residues 227–259. We predict that the amelogenin binding region is: (i) hydrophobic; (ii) largely disordered; and (iii) accessible to the external environment. In contrast, the binding region of CD63 is likely to be organized in a ‘7’ shape within the mushroom-like structure of CD63 EC2 (extracellular domain 2). In vivo, the protein interactions between the secreted enamel matrix proteins with the membrane-bound proteins are likely to occur at the specialized secretory surfaces of ameloblast cells called Tomes' processes. Such protein–protein interactions may be required to establish short-term order of the forming matrix and/or to mediate feedback signals to the transcriptional machinery of ameloblasts and/or to remove matrix protein debris during enamel biomineralization.

scholarly journals Lettuce necrotic yellows cytorhabdovirus protein localization and interaction map, and comparison with nucleorhabdoviruses

2012 ◽  
Vol 93 (4) ◽  
pp. 906-914 ◽  
Author(s):  
Kathleen M. Martin ◽  
Ralf G. Dietzgen ◽  
Renyuan Wang ◽  
Michael M. Goodin
Keyword(s):  
Protein Interactions ◽  
Matrix Protein ◽  
Fluorescent Protein ◽  
Protein Localization ◽  
Bimolecular Fluorescence Complementation ◽  
Cell Periphery ◽  
In Planta ◽  
Interaction Map ◽  
Yellow Dwarf Virus ◽  
P Proteins

Lettuce necrotic yellows virus (LNYV), Sonchus yellow net virus (SYNV) and Potato yellow dwarf virus (PYDV) are members of the family Rhabdoviridae that infect plants. LNYV is a cytorhabdovirus that replicates in the cytoplasm, while SYNV and PYDV are nucleorhabdoviruses that replicate in the nuclei of infected cells. LNYV and SYNV share a similar genome organization with a gene order of nucleoprotein (N), phosphoprotein (P), putative movement protein (Mv), matrix protein (M), glycoprotein (G) and polymerase (L). PYDV contains an additional predicted gene of unknown function located between N and P. In order to gain insight into the associations of viral structural and non-structural proteins and the mechanisms by which they may function, we constructed protein localization and interaction maps. Subcellular localization was determined by transiently expressing the viral proteins fused to green or red fluorescent protein in leaf epidermal cells of Nicotiana benthamiana. Protein interactions were tested in planta by using bimolecular fluorescence complementation. All three viruses showed Mv to be localized at the cell periphery and the G protein to be membrane associated. Comparing the interaction maps revealed that only the N–P and M–M interactions are common to all three viruses. Associations unique to only one virus include P–M for LNYV, G–Mv for SYNV and M–Mv, M–G and N–M for PYDV. The cognate N–P proteins of all three viruses interacted and exhibited characteristic changes in localization when co-expressed.

Co-evolution analysis to predict protein–protein interactions within influenza virus envelope

2014 ◽  
Vol 12 (02) ◽  
pp. 1441008 ◽  
Author(s):  
Ramil R. Mintaev ◽  
Andrei V. Alexeevski ◽  
Larisa V. Kordyukova
Keyword(s):  
Amino Acid ◽  
Protein Interactions ◽  
Influenza A ◽  
Matrix Protein ◽  
Amino Acid Sequences ◽  
Viral Envelope ◽  
Research Database ◽  
Virus Envelope ◽  
Physical Constraints ◽  
Matrix Protein M1

Interactions between integral membrane proteins hemagglutinin (HA), neuraminidase (NA), M2 and membrane-associated matrix protein M1 of influenza A virus are thought to be crucial for assembly of functionally competent virions. We hypothesized that the amino acid residues located at the interface of two different proteins are under physical constraints and thus probably co-evolve. To predict co-evolving residue pairs, the EvFold ( http://evfold.org ) program searching the (nontransitive) Direct Information scores was applied for large samplings of amino acid sequences from Influenza Research Database ( http://www.fludb.org/ ). Having focused on the HA, NA, and M2 cytoplasmic tails as well as C-terminal domain of M1 (being the less conserved among the protein domains) we captured six pairs of correlated positions. Among them, there were one, two, and three position pairs for HA–M2, HA–M1, and M2–M1 protein pairs, respectively. As expected, no co-varying positions were found for NA–HA, NA–M1, and NA–M2 pairs obviously due to high conservation of the NA cytoplasmic tail. The sum of frequencies calculated for two major amino acid patterns observed in pairs of correlated positions was up to 0.99 meaning their high to extreme evolutionary sustainability. Based on the predictions a hypothetical model of pair-wise protein interactions within the viral envelope was proposed.

Sign in / Sign up

Export Citation Format

Share Document