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91.
Rho kinase differentially regulates phosphorylation of nonmuscle myosin II isoforms A and B during cell rounding and migration 总被引:1,自引:0,他引:1
Sandquist JC Swenson KI Demali KA Burridge K Means AR 《The Journal of biological chemistry》2006,281(47):35873-35883
The actin-myosin cytoskeleton is generally accepted to produce the contractile forces necessary for cellular processes such as cell rounding and migration. All vertebrates examined to date are known to express at least two isoforms of non-muscle myosin II, referred to as myosin IIA and myosin IIB. Studies of myosin IIA and IIB in cultured cells and null mice suggest that these isoforms perform distinct functions. However, how each myosin II isoform contributes individually to all the cellular functions attributed to "myosin II" has yet to be fully characterized. Using isoform-specific small-interfering RNAs, we found that depletion of either isoform resulted in opposing migration phenotypes, with myosin IIA- and IIB-depleted cells exhibiting higher and lower wound healing migration rates, respectively. In addition, myosin IIA-depleted cells demonstrated impaired thrombin-induced cell rounding and undertook a more motile morphology, exhibiting decreased amounts of stress fibers and focal adhesions, with concomitant increases in cellular protrusions. Cells depleted of myosin IIB, however, were efficient in thrombin-induced cell rounding, displayed a more retractile phenotype, and maintained focal adhesions but only in the periphery. Last, we present evidence that Rho kinase preferentially regulates phosphorylation of the regulatory light chain associated with myosin IIA. Our data suggest that the myosin IIA and IIB isoforms are regulated by different signaling pathways to perform distinct cellular activities and that myosin IIA is preferentially required for Rho-mediated contractile functions. 相似文献
92.
93.
María Natalia Umaña Xiangcheng Mi Min Cao Brian J. Enquist Zhanqing Hao Robert Howe Yoshiko Iida Daniel Johnson Luxiang Lin Xiaojuan Liu Keping Ma I‐Fang Sun Jill Thompson Maria Uriarte Xugao Wang Amy Wolf Jie Yang Jess K. Zimmerman Nathan G. Swenson 《Global Ecology and Biogeography》2017,26(7):777-786
94.
Lora Swenson Ruth Green Rolf Joerger Michael Haas Karen Scott Yunyi Wei Urszula Derewenda David M. Lawson Zygmunt S. Derewenda 《Proteins》1994,18(3):301-306
A neutral lipase from the filamentous fungus Rhizopus delemar has been crystallized in both its proenzyme and mature forms. Although the latter crystallizes readily and produces a variety of crystal forms, only one was found to be suitable for X-ray studies. It is monoclinic (C2, a = 92.8 Å, b = 128.9 Å, c = 78.3 Å, β = 135.8) with two molecules in the asymmetric unit related by a noncrystallographic diad. The prolipase crystals are orthorhombic (P212121, with a = 79.8 Å, b = 115.2 Å, c = 73.0 Å) and also contain a pair of molecules in the asymmetric unit. Initial results of molecular replacement calculations using the refined coordinates of the related lipase from Rhizomucor miehei identified the correct orientations and positions of the protein molecules in the unit cells of crystals of both proenzyme and the mature form. © 1994 John Wiley & Sons, Inc. 相似文献
95.
We have quantitated the interactions of two rabbit skeletal troponin C fragments with troponin I and the troponin I inhibitory peptide. The calcium binding properties of the fragments and the ability of the fragments to exert control in the regulated actomyosin ATPase assay have also been studied. The N- and C-terminal divalent metal binding domains of rabbit skeletal troponin C, residues 1-97 and residues 98-159, respectively, were prepared by specific cleavage at cysteine-98 and separation by gel exclusion chromatography. Both of the troponin C fragments bind calcium. The calcium affinity of the weak sites within the N-terminal fragment is about an order of magnitude greater than is reported for these sites in troponin C, suggesting interaction between the calcium-saturated strong sites and the weak sites. Stoichiometric binding (1:1) of the troponin I inhibitory peptide to each fragment and to troponin C increased the calcium affinities of the fragments and troponin C. Complex formation was detected by fluorescence quenching or enhancement using dansyl-labeled troponin C (and fragments) or tryptophan-labeled troponin I inhibitory peptide. The troponin C fragments bind to troponin I with 1:1 stoichiometry and approximately equal affinities (1.6 x 10(6) M-1) which are decreased 4-fold in the presence of magnesium versus calcium. These calcium effects are much smaller than is observed for troponin C. The summed free energies for the binding of the troponin C fragments to troponin I are much larger than the free energy of binding troponin C. This suggests a large positive interaction free energy for troponin C binding to troponin I relative to the fragments.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
96.
Dana L. Swenson Kyoung-Jin Kim Erich W. Six Steven Clegg 《Molecular & general genetics : MGG》1994,244(2):216-218
The gene fimU, located on a recombinant plasmid carrying the Salmonella typhimurium type 1 fimbrial gene cluster is closely related to the Escherichia coli tRNA gene argU. The fimU gene complements an E. coli argU mutant that is a P2 lysogen, thereby allowing the phage P4 to grow in this strain but preventing the growth of phage lambda. In addition, fimU was shown to be involved in fimbrial expression since transformants of the E. coli argU mutant could produce fimbriae only in the presence of fimU but not in its absence, whereas in an E. coli argU
+ strain fimbriation did not require the fimU gene. 相似文献
97.
98.
The rotational relaxation times of nonpolymerizable skeletal and smooth muscle tropomyosin were measured by analysis of the decay of the zero-field birefringence at different temperatures and salt concentrations. Skeletal tropomyosin in solution is equally well modeled as a rigid rod or as a semiflexible rod with a persistence length of 150 nm. Smooth muscle tropomyosin does not fit the rigid rod model but is well approximated by a semiflexible rod model with a persistence length of 55 nm. The results indicate that smooth muscle tropomyosin is either a more flexible molecule than skeletal muscle tropomyosin or is a curved structure with an end-to-end length shorter than the coiled-coil contour length. Smooth muscle tropomyosin controls the actomyosin ATPase differently from skeletal muscle tropomyosin and it had been suggested that the reason is because it is more rigid; clearly, another explanation must be sought. 相似文献
99.
A technique for continuous intravenous infusion in cats 总被引:3,自引:0,他引:3
100.
The interaction between poly(α,L -lysine) (DP = 180) and glutaraldehyde was investigated in dilute aqueous solution by measurement of the kinetics of proton release at constant pH and temperature and at various concentrations of the reaction components. Under various conditions, the release of protons at constant pH appeared kinetically to be composed of at least two steps: an initial zero-order reaction, followed by a slower reaction. At excess of polylysine amino groups, the pH optimum for the rates of reaction was at pH 9–10 (24–25°C). Under the conditions used and at pH 8, the initial rate of the second kinetic step was proportional to the glutaraldehyde concentration and was practically independent of polylysine concentration at pH 8 and 8.6, at an excess of amino groups. At pH values of 7, 8, and 8.6 the apparent overall energy of activation for the second kinetic step was 18–19 kcal/mole (temp. range 4–40°C). Comparing acetaldehyde with the difunctional glutaraldehyde, it was found that the rate of proton release was much smaller in the case of acetaldehyde. Comparing n-butylamine with the macromolecular polylysine at equal concentrations of amino groups, the rates of proton release were much smaller in the case of n-butylamine. Collagen in aqueous medium also interacted with glutaraldehyde in a manner analogous to polylysine, although the conditions were not quite comparable. In the case of collagen, the initial fast proton liberation step was relatively much larger than in the case of polylysine. A reaction scheme for the initial reaction steps is being proposed which includes primary complex formation between glutaraldehyde and polylysine. This dialdehyde–polyamino acid system is considered to serve as a model for tanning processes of hides and for fixation procedures. 相似文献