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Italiano, J. E., Roberts, T. M., Stewart, M. and Fontana, C. A.
Fig. 6.

Comparison of the distribution of p48 and its phosphorylated form in detergent-permeabilized sperm by confocal fluorescence microscopy. Antiphosphotyrosine (green) labels the plasma membrane primarily at the leading edge of the lamellipod, whereas anti-p48 (red) labels the membrane uniformly. The green labeling in the cell body is due to recognition of fumarate reductase (see also Fig. 1 ) by antiphosphotyrosine. In this cell, anti-p48 failed to label the MOs. Bar, 10 μm.

Fig. 7.

Effect of pH modulation on phosphorylation of p48. Treatment of spermatozoa with acetate buffer at pH 5.5 (left panels) results in the disassembly of the MSP cytoskeleton. Labeling of the membrane with anti-p48 (red) is uniform but no antiphosphotyrosine labeling can be detected in the membrane. In cells fixed within 4 seconds after washing out the pH 5.5 buffer (middle panels), the cytoskeleton reforms along the lamellipod periphery and discrete spots of antiphosphotyrosine labeling appear along the membrane. By 60 seconds after acid removal (right panels) the cytoskeleton is completely reconstructed and spots of antiphosphotyrosine labeling are located along the lamellipodial leading edge. Bar, 10 μm. (B) Autoradiography of SDS-PAGE gels of S100 following incubation in P-γ-ATP at pH 5.5 vs pH 7.0. Labeling of p48 (arrow) is undetectable at the acidic pH.

The fluorescence labeling pattern observed in response to alteration of intracellular pH correlates with the pH sensitivity of labeling of p48 with 32 P-γ-ATP in S100. As shown in Fig. 7B , phosphorylation of p48 is readily detectable in S100 at pH 7. By contrast, when the pH of S100 was lowered to 5.5 before addition of 32 P-γ-ATP, labeling of p48 was barely detectable.

To examine the pattern of phosphorylation of p48, we incubated vesicles isolated from S100 with 32 P-γ-ATP under a range of different conditions and monitored phosphorylation by autoradiography. As shown in Womens Oklahoma City Thunder Cashmere Reversible Scarf The Elder Statesman X NBA 0ELayUz
, when membrane vesicles alone were incubated with 32 P-γ-ATP for 30 minutes, labeling of p48 was barely detectable. However, adding cytosol to these vesicles resulted in rapid phosphorylation of p48 so that within 5 minutes the protein became heavily labeled. No significant protein phosphorylation was detected when 32 P-γ-ATP was incubated with cytosol. Therefore, it appears that phosphorylation of p48 requires a cytosolic factor.

Fig. 8.

Phosphorylation of p48 requires cytosolic components. Coomassie-stained SDS-PAGE gels (left) and autoradiograms (center) of equal aliquots of vesicles incubated with P-γ-ATP for the intervals indicated. Labeling of p48 (arrow) is barely detectable after 30 minutes incubation of vesicles alone but increases dramatically by 5 minutes after addition of cytosol. Incubation of cytosol alone in P-γ-ATP for 5 minutes yields almost no protein labeling.

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Reconstitution of the MSP-based motility system from Ascaris sperm indicated that the lamellipodial membrane has an important role in directing the spatial distribution of cytoskeletal assembly fundamental to locomotion ( Italiano et al., 1996 ). Here, we have tested this hypothesis by fractionating sperm membrane proteins and reconstituting MSP polymerization in vitro and thus have shown that a 48 kDa integral membrane phosphoprotein is required for the localized membrane-associated polymerization of MSP. This protein is found exclusively in the vesicle fraction of the cell-free extract (S100) in which motility is reconstituted and, when extracted from the bilayer, purified and recombined with cytosol, it induces the assembly of MSP filaments. The activity of p48 is regulated by tyrosine phosphorylation, which appears to be catalyzed by a cytosolic tyrosine kinase.

First electronic television system on 180 lines at 25 fps was created in the beginning of 1935 in Leningrad (St. Petersburg). In September 1937 the experimental Leningrad TV Center (OLTC) was put in action. OLTC worked with 240 lines at 25 fps progressive scan. Handkerchief Dark blue with a meandering gold pattern Notch Vi2rzbjxu

In Moscow, experimental transmissions of electronic television took place on March 9, 1937, using equipment manufactured by . Regular broadcasting began on December 31, 1938. It was quickly realized that 343 lines of resolution offered by this format would have become insufficient in the long run, thus a specification for 441-line format at 25 fps interlaced was developed in 1940. [144]

Television broadcasts were suspended during Great Patriotic War . In 1944, while the war was still raging, a new standard, offering 625 lines of vertical resolution was prepared. This format was ultimately accepted as a national standard. [144]

The transmissions in 625-line format started in Moscow on November 4, 1948. Regular broadcasting began on June 16, 1949. Details for this standard were formalized in 1955 specification called GOST 7845-55, basic parameters for black-and-white television broadcast . In particular, frame size was set to 625 lines, frame rate to 25 frames/s interlaced, and video bandwidth to 6MHz. These basic parameters were accepted by most countries having 50Hz mains frequency and became the foundation of television systems presently known as PAL and SECAM.

Starting in 1951, broadcasting in the 625-line standard was introduced in other major cities of the Soviet Union.

Color television broadcast started in 1967, using SECAM color system. [91]

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