5.3.1 Infection of cells
Cells were seeded one day or few hours before infection. One colony of pathogenic S.aureus is grown in 1ml TSB medium overnight at 37°C. Prior to infection, bacteria are diluted 1:10 in fresh TSB medium and incubated for another 2 h to obtain bacteria in the logarithmic growth state minimizing toxin production.
After culture, bacteria are washed twice with PBS. For some experiments, bacteria are incubated with fluorescent dyes or fibronectin for 30 min to 1h at 37°C. Following this incubation, bacteria are washed three times again in PBS. The number of bacteria is photometrically determined at OD600 using a standard curve. Cells are infected at proper MOIs (for FACS or microscopic analysis: MOI 30, for gentamicin protection assay: MOI 20) depending on the experiment.
5.3.2 Gentamicin Protection Assay
2x105 HEK293T cells or 5x104 MEFs or 5x104 HeLa cells are seeded in 10μg/ml poly-lysine coated 24-well plates and infected at MOI 20 for 2 h at 37°C and 5% CO2. 2h later, to evaluate the number of intracellular bacteria, the medium is carefully removed and replaced by fresh medium containing 50 μg/ml gentamicin. After incubation for 1 h at 37°C, intracellular bacteria are released by treatment of 0.5% saponin for 15 min at 37°C. Samples are gradiently diluted (for S.aureus, first dilution: 100μl bacteria suspension in 900μl PBS; second dilution: 100μl of first dilution in 900μl PBS) and
General Material and Methods
114
20μl suspension from second dilution is plated on TSB agar plates to determine the recovered colony forming units (cfu). Simultaneously, the parallel infected cells are directly lysed with 0.5% saponin without Gentamicin treatment and gradiently diluted (for S.aureus, first dilution: 40μl bacteria suspension in 960μl PBS; second dilution:
100μl of first dilution in 900μl PBS) and 20μl suspension from second dilution is plated on TSB agar plates to quantify cell adherent bacteria.
5.3.3 Extra- and intracellular bacteria staining and fluorescence microscopy examination
For PIP5KIγ90–/–and control MEF cells, 3 × 104 cells for each cell line are seeded on 10μg/ml poly-L-lysine coated, acid-washed glass cover slips in 24-well plate. Next day, cells are infected with S.aureus at MOI 30 for 2 h, then washed twice with PBS+/+ and fixed with 4% PFA for 20 min at RT. After this, cells are incubated in blocking buffer (PBS+/+ plus 10% FCS) for 10 min. Extracellular bacteria are detected by rabbit polyclonal α-staphylococcal serum diluted in blocking buffer (45 min at RT).
Afterwards, samples are washed three times with PBS+/+ and incubated with goat α-rabbit IgG-Cy5 in the dark for 30 min. Then, cells are washed three times and permeabilized by 0.5% Triton/PBS. 10 min later, cells are washed three times with PBS+/+, blocked for 10 min, and incubated with rabbit polyclonal α-staphylococcal serum at RT. 45min later, samples are washed three times with PBS+/+ and incubated with goat α-rabbit IgG coupled to Cy2 in the dark for 30 min. Finally, after three washes with PBS+/+, the cover slips are embedded in mounting medium (DaKo, Glostrup, Denmark) on glass slides and sealed with nail polish. Images are acquired with Leica
General Material and Methods
115
AF6000LX fluorescence microscope and processed with ImageJ.
For the experiment performed with HEK293T cells, 2 x 105 transfected 293cells are seeded on poly-L-lysine coated cover slips in 24-well plate. 2 hours later, cells are infected with pacific-blue stained and biotin-labeled S.aureus at MOI 30 for 2 h, then washed twice with PBS+/+ and fixed with 4% PFA for 20 min at RT. After this, cells are incubated in blocking solution for 10min, afterwards, cells are incubated with streptavidin-AlexaFluor647 for 1h at RT in the dark. Finally, after three washes with PBS+/+, the cover slips are transferred to glass slides, embedded in mounting medium and sealed with nail polish. Images were acquired with Leica AF6000LX fluorescence microscope.
For immunofluorescence staining of transfected PIP5KIγ90–/– orNIH3T3 SYNJ1 KO cells, 5 × 104 cells are seeded on poly-L-lysine coated cover slips in 24-well plate. Next day, cells are transfected with indicated plasmid DNA. 24h or 48h after transfection, cells were infected with pacific-blue stained and biotin-labeled S.aureus at MOI 30 for 2 h. After the infection, the same staining procedure as described above for the transfected 293 cells was used.
5.3.4 Invasion assay by flow cytometry
This method is based on the paper from Hauck lab (Pils, Stefan, et al. 2006). 1x106 HEK293T cells or 2 x105 NIH3T3 cells are seeded in poly-lysine coated wells of 6-well plate. One colony of S.aureus is cultured in 1ml TSB medium overnight. Next day, Bacterial culture is transferred to 9ml fresh TSB medium and cultured for another 2h.
Afterwards, S.aureus is stained with FITC and the OD600 is measured. Cells are
General Material and Methods
116
infected at MOI30 and incubated for 2h at 37°C. After 2h infection, medium is aspirated and cells are washed once with PBS. 1ml Trypsin/EDTA is added to each well and incubated for 2min at 37°C. 3ml fresh cell culture medium is added to stop trypsinization and cells are separated via pipetting up and down several times. Then, cells are transferred into 15ml tubes and centrifuged. The cell pellet is re-suspended in 1ml ice cold FACS buffer and transferred to 1.5 ml Eppendorf tubes, then washed twice again. The cells are re-suspended in 1ml FACS buffer and distributed to 2 FACS tubes (one tube 600μl and the other 400μl) and placed on ice, under dark. The samples containing 600μl cell suspension are measured by Flow cytometry. To quench the fluorescence from adhering bacteria on cell surface, 400μl trypan blue is added and the signal is measured again. The fluorescence intensity is analyzed to compare the invasion efficiency.
5.3.5 live-cell imaging
Cells are seeded in poly-lysine coated 3.5cm dishes (with integrated coverslip), eg. 1 x 105 cells/dish. Next day, exchange of DMEM medium with colorless DMEM and preparation of the CLSM (heating of incubation chamber to 37°C and connection of CO2 supply) followed. If bacterial infection is investigated, cells are directly infected with the fluorescence stained S.aureus at MOI60. First of all, appropriate cells are searched (for example, the transfected cells). Then the settings are adjusted and finally the movie is started to record. Afterwards, images and movies are analyzed by LAS software and Image J.
5.3.6 Scanning-Electron-Microscopy (SEM)
General Material and Methods
117
Cells are seeded on poly-lysine coated coverslips in a 24-well plate (3 x 104 cells/well).
Next day, cells are infected at MOI 50. Subsequently cells are fixed with SEM fixation solution for 1h at RT. Afterwards samples are gradually dehydrated via an ethanol dilution range. The critical point drying by liquid CO2 followed and samples are sputtered with 10 nm gold-palladium. Samples are analyzed by a Philips SEM 505.
5.3.7 Measurement of FAK phosphorylation upon S.aureus infection
Cells are starved in starvation medium (DMEM plus 0.5% FCS) overnight. Next day, cells are detached by Trypsin/EDTA and the trypsinization is stopped via the addition of trypsin inhibitor. After centrifugation, cells are re-suspended in suspension medium (DMEM plus 0.25% BAS) and further incubated for 45min at 37°C. Meanwhile, 6 cm dishes are coated with poly-lysine for 45min and later blocked by suspension medium for another 45min, afterwards, 2 x 106 cells are seeded in each coated dish and incubated for 45min to let cells attach. Then cells are infected with 2μg/ml Fn coated bacteria at MOI 100for indicated time. Whole cell lysate is prepared with the modified RIPA buffer and FAK phosphorylation is analyzed via western blotting.
References
118
References
Agerer, Franziska, et al. "Cellular invasion by Staphylococcus aureus reveals a functional link between focal adhesion kinase and cortactin in integrin-mediated internalisation." Journal of cell science 118.10 (2005): 2189-2200.
Agerer, Franziska, et al. "Integrin-mediated invasion of Staphylococcus aureus into human cells requires Src family protein-tyrosine kinases." Journal of Biological Chemistry 278.43 (2003): 42524-42531.
Alva-Murillo, Nayeli, Joel Edmundo López-Meza, and Alejandra Ochoa-Zarzosa.
"Nonprofessional Phagocytic Cell Receptors Involved in Staphylococcus aureus Internalization." BioMed research international 2014 (2014).
Antonescu, Costin N., et al. "Phosphatidylinositol-(4, 5)-bisphosphate regulates clathrin-coated pit initiation, stabilization, and size." Molecular biology of the cell 22.14 (2011): 2588-2600.
Attree, Olivier, et al. "The Lowe's oculocerebrorenal syndrome gene encodes a protein highly homologous to inositol polyphosphate-5-phosphatase." (1992): 239-242.
Balla, Tamas, and Péter Várnai. "Visualization of Cellular Phosphoinositide Pools with GFP‐Fused Protein‐Domains." Current Protocols in Cell Biology (2009): 24-4.
Balla, Tamas. "Phosphoinositides: tiny lipids with giant impact on cell regulation."
Physiological reviews 93.3 (2013): 1019-1137.
Banno, Asoka, and Mark H. Ginsberg. "Integrin activation." Biochemical Society Transactions 36.Pt 2 (2008): 229.
Barczyk, Malgorzata, Sergio Carracedo, and Donald Gullberg. "Integrins." Cell and tissue research 339.1 (2010): 269-280.
Bayles, Kenneth W., et al. "Intracellular staphylococcus aureusescapes the endosome and induces apoptosis in epithelial cells." Infection and immunity 66.1 (1998): 336-342.
Borisova, Marina, et al. "Integrin-mediated internalization of Staphylococcus aureus does not require vinculin." BMC cell biology 14.1 (2013): 2.
Boronenkov, Igor V., et al. "Phosphoinositide signaling pathways in nuclei are associated with nuclear speckles containing pre-mRNA processing factors." Molecular biology of the cell 9.12 (1998): 3547-3560.
References
119
Bost, Kenneth L., et al. "Staphylococcus aureus infection of mouse or human osteoblasts induces high levels of interleukin-6 and interleukin-12 production." Journal of Infectious Diseases 180.6 (1999): 1912-1920.
Böttcher, Ralph Thomas, et al. "Sorting nexin 17 prevents lysosomal degradation of [beta] 1 integrins by binding to the [beta] 1-integrin tail." Nature cell biology 14.6 (2012): 584-592.
Boucrot, Emmanuel, et al. "Role of lipids and actin in the formation of clathrin-coated pits." Experimental cell research 312.20 (2006): 4036-4048.
Bouvard, Daniel, et al. "Integrin inactivators: balancing cellular functions in vitro and in vivo." Nature Reviews Molecular Cell Biology 14.7 (2013): 430-442.
Brakebusch, Cord, and Reinhard Fässler. "The integrin–actin connection, an eternal love affair." The EMBO journal 22.10 (2003): 2324-2333.
Brose, Nils, Andrea Betz, and Heike Wegmeyer. "Divergent and convergent signaling by the diacylglycerol second messenger pathway in mammals." Current opinion in neurobiology 14.3 (2004): 328-340.
Bur, Stephanie, et al. "The Staphylococcus aureus extracellular adherence protein promotes bacterial internalization by keratinocytes independent of fibronectin-binding proteins." Journal of Investigative Dermatology 133.8 (2013): 2004-2012.
Burke, Fiona M., et al. "Fibronectin-binding protein B variation in Staphylococcus aureus." BMC microbiology 10.1 (2010): 160.
Calderwood, David A. "Integrin activation." Journal of cell science 117.5 (2004): 657-666.
Campbell, Iain D., and Martin J. Humphries. "Integrin structure, activation, and interactions." Cold Spring Harbor perspectives in biology 3.3 (2011): a004994.
Campellone, Kenneth G., and Matthew D. Welch. "A nucleator arms race: cellular control of actin assembly." Nature reviews Molecular cell biology 11.4 (2010): 237-251.
Chambers, Henry F., and Frank R. DeLeo. "Waves of resistance: Staphylococcus aureus in the antibiotic era." Nature Reviews Microbiology 7.9 (2009): 629-641.
Chang-Ileto, Belle, et al. "Synaptojanin 1-mediated PI (4, 5) P 2 hydrolysis is modulated
References
120
by membrane curvature and facilitates membrane fission." Developmental cell 20.2 (2011): 206-218.
Chinthalapudi, Krishna, et al. "Lipid binding promotes oligomerization and focal adhesion activity of vinculin." The Journal of cell biology 207.5 (2014): 643-656.
Cossart, Pascale, and Philippe J. Sansonetti. "Bacterial invasion: the paradigms of enteroinvasive pathogens." Science 304.5668 (2004): 242-248.
Cullen, Peter J. "Endosomal sorting and signalling: an emerging role for sorting nexins." Nature reviews Molecular cell biology 9.7 (2008): 574-582.
Cullen, Peter J., et al. "Modular phosphoinositide-binding domains–their role in signalling and membrane trafficking." Current Biology 11.21 (2001): R882-R893.
Czech, Michael P. "PIP2 and PIP3: complex roles at the cell surface." Cell 100.6 (2000):
603-606.
Da Silva, Claudio Vieira, et al. "A glance at Listeria and Salmonella cell invasion:
different strategies to promote host actin polymerization." International Journal of Medical Microbiology 302.1 (2012): 19-32.
Dehio, Christoph, et al. "Interaction of Bartonella henselae with endothelial cells results in bacterial aggregation on the cell surface and the subsequent engulfment and internalisation of the bacterial aggregate by a unique structure, the invasome." Journal of Cell Science 110.18 (1997): 2141-2154.
Deitch, Edwin A., et al. "Caco-2 and IEC-18 intestinal epithelial cells exert bactericidal activity through an oxidant-dependent pathway." Shock 4.5 (1995): 345-350.
Delon, Isabelle, and Nicholas H. Brown. "Integrins and the actin cytoskeleton." Current opinion in cell biology 19.1 (2007): 43-50.
Di Paolo, Gilbert, and Pietro De Camilli. "Phosphoinositides in cell regulation and membrane dynamics." Nature 443.7112 (2006): 651-657.
Di Paolo, Gilbert, et al. "Recruitment and regulation of phosphatidylinositol phosphate kinase type 1γ by the FERM domain of talin." Nature 420.6911 (2002): 85-89.
Drouet, Valérie, and Suzanne Lesage. "Synaptojanin 1 Mutation in Parkinson’s Disease Brings Further Insight into the Neuropathological Mechanisms." BioMed research international 2014 (2014).
References
121
Dziewanowska, Katarzyna, et al. "Staphylococcal fibronectin binding protein interacts with heat shock protein 60 and integrins: role in internalization by epithelial cells."
Infection and immunity 68.11 (2000): 6321-6328.
Elliott, Paul R., et al. "The Structure of the talin head reveals a novel extended conformation of the FERM domain." Structure 18.10 (2010): 1289-1299.
Erdmann, Kai S., et al. "A role of the Lowe syndrome protein OCRL in early steps of the endocytic pathway." Developmental cell 13.3 (2007): 377-390.
Falkenburger, Björn H., Jill B. Jensen, and Bertil Hille. "Kinetics of M1 muscarinic receptor and G protein signaling to phospholipase C in living cells." The Journal of general physiology 135.2 (2010): 81-97.
Foster, Timothy J., et al. "Adhesion, invasion and evasion: the many functions of the surface proteins of Staphylococcus aureus." Nature Reviews Microbiology 12.1 (2014):
49-62.
Fowler, Trent, et al. "Cellular invasion by Staphylococcus aureus involves a fibronectin bridge between the bacterial fibronectin-binding MSCRAMMs and host cell β1 integrins." European journal of cell biology 79.10 (2000): 672-679.
Fowler, Trent, et al. "Src kinase has a central role in in vitro cellular internalization of Staphylococcus aureus." Cellular microbiology 5.6 (2003): 417-426.
Fraunholz, Martin, and Bhanu Sinha. "Intracellular Staphylococcus aureus: live-in and let die." The Staphylococci and staphylococcal pathogenesis (2012): 116.
Fukami, Kiyoko, et al. "alpha-Actinin and vinculin are PIP2-binding proteins involved in signaling by tyrosine kinase." Journal of Biological Chemistry 269.2 (1994): 1518-1522.
Garzoni, Christian, and William L. Kelley. "Staphylococcus aureus: new evidence for intracellular persistence." Trends in microbiology 17.2 (2009): 59-65.
Garzoni, Christian, et al. "A global view of Staphylococcus aureus whole genome expression upon internalization in human epithelial cells." BMC genomics 8.1 (2007):
171.
Geiger, Benjamin, and Kenneth M. Yamada. "Molecular architecture and function of matrix adhesions." Cold Spring Harbor perspectives in biology 3.5 (2011): a005033.
References
122
Geiger, Benjamin, Joachim P. Spatz, and Alexander D. Bershadsky. "Environmental sensing through focal adhesions." Nature reviews Molecular cell biology 10.1 (2009):
21-33.
Geoghegan, Joan A., et al. "Subdomains N2N3 of fibronectin binding protein A mediate Staphylococcus aureus biofilm formation and adherence to fibrinogen using distinct mechanisms." Journal of bacteriology 195.11 (2013): 2675-2683.
Giancotti, Filippo G., and Erkki Ruoslahti. "Integrin signaling." Science 285.5430 (1999): 1028-1033.
Goksoy, Esen, et al. "Structural basis for the autoinhibition of talin in regulating integrin activation." Molecular cell 31.1 (2008): 124-133.
Goldschmidt-Clermont, Pascal J., et al. "The actin-binding protein profilin binds to PIP2 and inhibits its hydrolysis by phospholipase C." Science 247.4950 (1990): 1575-1578.
Goñi, Guillermina M., et al. "Phosphatidylinositol 4, 5-bisphosphate triggers activation of focal adhesion kinase by inducing clustering and conformational changes."
Proceedings of the National Academy of Sciences 111.31 (2014): E3177-E3186.
Götz, Friedrich, Tammy Bannerman, and Karl-Heinz Schleifer. "The genera staphylococcus and macrococcus." The prokaryotes. Springer US, 2006. 5-75.
Goult, Benjamin T., et al. "Structural studies on full-length talin1 reveal a compact auto-inhibited dimer: implications for talin activation." Journal of structural biology 184.1 (2013): 21-32.
Haffner, Christof, et al. "Direct interaction of the 170 kDa isoform of synaptojanin 1 with clathrin and with the clathrin adaptor AP-2." Current Biology 10.8 (2000): 471-474.
Haggar, Axana, et al. "Extracellular adherence protein from Staphylococcus aureus enhances internalization into eukaryotic cells." Infection and immunity 71.5 (2003):
2310-2317.
Haglund, Cat M., and Matthew D. Welch. "Pathogens and polymers: microbe–host interactions illuminate the cytoskeleton." The Journal of cell biology 195.1 (2011): 7-17.
References
123
Ham, Hyeilin, Anju Sreelatha, and Kim Orth. "Manipulation of host membranes by bacterial effectors." Nature Reviews Microbiology 9.9 (2011).
Harburger, David S., and David A. Calderwood. "Integrin signalling at a glance."
Journal of cell science 122.2 (2009): 159-163.
Hauck, Christof R. "Cell adhesion receptors–signaling capacity and exploitation by bacterial pathogens." Medical microbiology and immunology 191.2 (2002): 55-62.
Hauck, Christof R., and Knut Ohlsen. "Sticky connections: extracellular matrix protein recognition and integrin-mediated cellular invasion by Staphylococcus aureus." Current opinion in microbiology 9.1 (2006): 5-11.
Hauck, Christof R., Marina Borisova, and Petra Muenzner. "Exploitation of integrin function by pathogenic microbes." Current opinion in cell biology 24.5 (2012): 637-644.
Hennekinne, Jacques-Antoine, et al. "How should staphylococcal food poisoning outbreaks be characterized?." Toxins 2.8 (2010): 2106-2116.
Higgs, Henry N., and Thomas D. Pollard. "Activation by Cdc42 and PIP2 of Wiskott-Aldrich syndrome protein (WASp) stimulates actin nucleation by Arp2/3 complex."
The Journal of cell biology 150.6 (2000): 1311-1320.
Hiramatsu, Keiichi. "Vancomycin-resistant Staphylococcus aureus: a new model of antibiotic resistance." The Lancet infectious diseases 1.3 (2001): 147-155.
Hirschhausen, Nina, et al. "A novel staphylococcal internalization mechanism involves the major autolysin Atl and heat shock cognate protein Hsc70 as host cell receptor."
Cellular microbiology 12.12 (2010): 1746-1764.
Hoffmann, Christine, et al. "Caveolin limits membrane microdomain mobility and integrin-mediated uptake of fibronectin-binding pathogens." Journal of cell science 123.24 (2010): 4280-4291.
Hoffmann, Christine, Knut Ohlsen, and Christof R. Hauck. "Integrin-mediated uptake of fibronectin-binding bacteria." European journal of cell biology 90.11 (2011): 891-896.
Holland, Thomas L., Christopher Arnold, and Vance G. Fowler. "Clinical management of Staphylococcus aureus bacteremia: a review." Jama 312.13 (2014): 1330-1341.
References
124
Hopper, Neil A., and Vincent O'Connor. "Ephrin tempers two-faced synaptojanin 1."
Nature cell biology 7.5 (2005): 454-456.
Hsu, FoSheng, and Yuxin Mao. "The structure of phosphoinositide phosphatases:
Insights into substrate specificity and catalysis." Biochimica et Biophysica Acta (BBA)-Molecular and Cell Biology of Lipids 1851.6 (2015): 698-710.
Hudson, Michael C., et al. "Internalization of Staphylococcus aureus by cultured osteoblasts." Microbial pathogenesis 19.6 (1995): 409-419.
Hughes, Diarmaid. "Exploiting genomics, genetics and chemistry to combat antibiotic resistance." Nature reviews genetics 4.6 (2003): 432-441.
Humphries, Jonathan D., Adam Byron, and Martin J. Humphries. "Integrin ligands at a glance." Journal of cell science 119.19 (2006): 3901-3903.
Huttenlocher, Anna, and Alan Rick Horwitz. "Integrins in cell migration." Cold Spring Harbor perspectives in biology 3.9 (2011): a005074.
Huveneers, Stephan, and Erik HJ Danen. "Adhesion signaling–crosstalk between integrins, Src and Rho." Journal of cell science 122.8 (2009): 1059-1069.
Hynes, Richard O. "Integrins: bidirectional, allosteric signaling machines." Cell 110.6 (2002): 673-687.
Irie, Fumitoshi, et al. "EphrinB–EphB signalling regulates clathrin-mediated endocytosis through tyrosine phosphorylation of synaptojanin 1." Nature cell biology 7.5 (2005): 501-509.
Isberg, Ralph R., and Guy Tran Van Nhieu. "The mechanism of phagocytic uptake promoted by invasin-integrin interaction." Trends in cell biology 5.3 (1995): 120-124.
Ishihara, Hisamitsu, et al. "Cloning of cdnas encoding two isoforms of 68-kda type i phosphatidylinositol4-phosphate 5-kinase." Journal of Biological Chemistry 271.39 (1996): 23611-23614.
Ishihara, Hisamitsu, et al. "Type I Phosphatidylinositol-4-phosphate 5-Kinases cloning of the third isoform and deletion/substitution analysis of members of this novel lipid kinase family." Journal of Biological Chemistry 273.15 (1998): 8741-8748.
Ivaska, Johanna. "Unanchoring integrins in focal adhesions." Nature cell biology 14.10 (2012): 981-983.
References
125
Janmey, Paul A. "Phosphoinositides and calcium as regulators of cellular actin assembly and disassembly." Annual Review of Physiology 56.1 (1994): 169-191.
Janmey, Paul A., and Uno Lindberg. "Cytoskeletal regulation: rich in lipids." Nature Reviews Molecular Cell Biology 5.8 (2004): 658-666.
Jänne, Pasi A., et al. "Functional overlap between murine Inpp5b and Ocrl1 may explain why deficiency of the murine ortholog for OCRL1 does not cause Lowe syndrome in mice." Journal of Clinical Investigation 101.10 (1998): 2042.
Johnson, Jason M., et al. "Genome-wide survey of human alternative pre-mRNA splicing with exon junction microarrays." Science 302.5653 (2003): 2141-2144.
Jones, David R., et al. "Nuclear PtdIns5P as a transducer of stress signaling: an in vivo role for PIP4Kbeta." Molecular cell 23.5 (2006): 685-695.
Kadamur, Ganesh, and Elliott M. Ross. "Mammalian phospholipase C." Annual review of physiology 75 (2013): 127-154.
Kharitidi, Dmitri, et al. "Interplay of Endosomal pH and Ligand Occupancy in Integrin α5β1 Ubiquitination, Endocytic Sorting, and Cell Migration." Cell reports 13.3 (2015):
599-609.
Kirkbride, Kellye C., et al. "Cortactin: a multifunctional regulator of cellular invasiveness." Cell adhesion & migration 5.2 (2011): 187-198.
Kong, Xiangming, et al. "Structural basis for the phosphorylation-regulated focal adhesion targeting of type Iγ phosphatidylinositol phosphate kinase (PIPKIγ) by talin."
Journal of molecular biology 359.1 (2006): 47-54.
Krauß, Michael, and Volker Haucke. "Phosphoinositides: regulators of membrane traffic and protein function." FEBS letters 581.11 (2007): 2105-2111.
Kühbacher, Andreas, et al. "Phosphatidylinositol 5-phosphatase oculocerebrorenal syndrome of Lowe protein (OCRL) controls actin dynamics during early steps of Listeria monocytogenes infection." Journal of Biological Chemistry 287.16 (2012):
13128-13136.
Kutateladze, Tatiana G. "Phosphatidylinositol 3-phosphate recognition and membrane docking by the FYVE domain." Biochimica et Biophysica Acta (BBA)-Molecular and Cell Biology of Lipids 1761.8 (2006): 868-877.
References
126
Larson, Richard S., et al. "Primary structure of the leukocyte function-associated molecule-1 alpha subunit: an integrin with an embedded domain defining a protein superfamily." The Journal of cell biology 108.2 (1989): 703-712.
Le, Oanh Thi Tu, et al. "Phosphorylation of phosphatidylinositol 4-phosphate 5-kinase γ by Akt regulates its interaction with talin and focal adhesion dynamics." Biochimica
Le, Oanh Thi Tu, et al. "Phosphorylation of phosphatidylinositol 4-phosphate 5-kinase γ by Akt regulates its interaction with talin and focal adhesion dynamics." Biochimica