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91.
Boguslaw Szewczyk Krystyna Bienkowska-Szewczyk Lloyd M. Kozloff 《Molecular & general genetics : MGG》1986,202(3):363-367
Summary The proteins synthesized in Escherichia coli B cells after infection with various T4 bacteriophage tail baseplate mutants were analysed by the immunoblotting method for the presence of the 15 Kilodalton lysozyme found in phage T4 particles. Using three different antisera: anti-phage, anti-baseplate and anti-15K lysozyme, it has been found that the 15K lysozyme is not present in lysates of bacteria infected with T4 gene 25 amber mutants. The 15K lysozyme was also found to be expressed in E. coli B cells transformed with a plasmid containing only a small portion of the T4 genome but which included T4 gene 25. These observations indicate that the 15K lysozyme is the gene 25 product. 相似文献
92.
Protein E1, a porin of the outer membrane of Pseudomonas aeruginosa, was reconstituted into planar lipid bilayers. Single channel conductance of the protein appeared to be 230 pS (pico siemens) in 1 M KCl-10 mM Hepes, pH7.2. This value is approximately 5 times lower than the conductance of the OmpF channel of Escherichia coli. Conductance increased linearly as the membrane potential was raised from -200 mV to +200 mV, and was nearly proportional to the KCl concentration. These results show that protein E1 is probably a genuine porin in the P. aeruginosa outer membrane supporting the earlier conclusion that protein E1 forms a small channel. 相似文献
93.
Pulmonary lesions seen in autopsies of leprosy patients were initially thought to involve microsporidial infection. After immunohistochemical studies, it was concluded that the infectious microorganism was Cryptococcus neoformans. 相似文献
94.
K Mishima M Terashima S Obara K Yamada K Imai M Shimoyama 《Journal of biochemistry》1991,110(3):388-394
We have reported the purification and characterization of arginine-specific ADP-ribosyltransferase from hen liver nuclei [Tanigawa, Y. et al. (1984) J. Biol. Chem. 259, 2022-2029] and the DNA-dependent mono(ADP-ribosyl)ation of p33, an acceptor protein in the nuclei [Mishima, K. et al. (1989) Eur. J. Biochem. 179, 267-273]. In the present study, we obtained evidence that among various tissues and cells from chicken, polymorphonuclear cells, so-called heterophils, possess both the ADP-ribosyltransferase and p33 at high levels. Percoll density gradient centrifugation of the postnuclear fraction of the heterophils revealed the co-localization of ADP-ribosyltransferase with p33 in the granule fraction. The enzyme and p33 were purified approximately 219- and 3.77-fold, respectively, from postnuclear pellet fraction to apparent homogeneity. The properties of heterophil ADP-ribosyltransferase and p33 were compared with those of the liver enzyme and p33. The molecular mass of the heterophil enzyme was estimated by SDS-polyacrylamide gel electrophoresis to be 27.5 kDa. The enzyme activity was stimulated by a sulfhydryl agent and inhibited by lysolecithin, NaCl, and inorganic phosphate. The mono(ADP-ribosyl)ation of p33 was markedly enhanced by polyanion, such as DNA, RNA, or poly(L-glutamate). SDS-polyacrylamide gel electrophoretic analysis after limited trypsin proteolysis of p33s, purified from chicken heterophils and liver, showed much the same pattern. Thus, it appears that ADP-ribosyltransferase and p33 present in heterophils are identical to those in the liver, respectively. p33 is considered to be an in situ substrate for ADP-ribosyltransferase, since it was specifically mono(ADP-ribosyl)ated in permeabilized heterophils.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
95.
R D Bowditch C E Halloran S Aota M Obara E F Plow K M Yamada M H Ginsberg 《The Journal of biological chemistry》1991,266(34):23323-23328
The binding of fibronectin (Fn) to several integrins involves the Arg-Gly-Asp (RGD) tripeptide sequence. However, linear synthetic RGD peptides do not completely mimic the cell attachment activity of intact Fn or certain large Fn fragments. This suggests that the integrin-Fn interaction involves a more extended surface of Fn than that provided by the RGD sequence. To test this possibility, three novel monoclonal anti-Fn antibodies that inhibit its binding to a purified integrin, alpha IIb beta 3, were developed. The epitopes of these three antibodies mapped to a region at least 55 residues amino-terminal of the RGD sequence. Further, recombinant fragments of Fn containing these epitopes and lacking the RGD site also inhibited the binding of Fn to purified alpha IIb beta 3. These fragments, which spanned Fn residues 1359-1436, bound to alpha IIb beta 3 in a divalent cation-dependent manner. In addition, this region of Fn bound specifically to alpha IIb beta 3 on thrombin-stimulated but not resting platelets. These results demonstrate the presence of additional sequences in Fn that interact with integrin alpha IIb beta 3 and suggest that multiple sites in Fn are involved in its recognition by this integrin. 相似文献
96.
Andreina Poggi Boguslaw Rucinski Pranee James John C. Holt Stefan Niewiarowski 《Experimental cell research》1984,150(2)
Platelet derived growth factor (PDGF) has been partially purified from porcine platelets. Purification steps included heparin-agarose chromatography of the material released by thrombin-stimulated washed porcine platelets and Blue-Sepharose chromatography. Preparative isoelectric focusing showed that isoelectric point of porcine PDGF is at pH 10.0–11.0 and elution experiments from sodium dodecyl sulfate (SDS) polyacrymlam de gels indicated that its molecular weight is close to 30 kD. The immunoglobulin fraction prepared from anti-human PDGF serum inhibited the mitogenic activity of porcine PDGF. These experiments suggest a homology of porcine and human PDGF. Porcine platelet factor 4 and porcine platelet basic protein were devoid of significant mitogenic activity. 相似文献
97.
Site-directed mutagenesis of the cell-binding domain of human fibronectin: separable, synergistic sites mediate adhesive function 总被引:40,自引:0,他引:40
Polypeptide sequences required for function of the cell-binding domain of human fibronectin were analyzed by site-directed mutagenesis. Site-specific deletion of the putative recognition sequence Arg-Gly-Asp-Ser or an Asp-to-Glu mutation decreased the adhesive activity of fibronectin fusion proteins expressed in E. coli by greater than or equal to 97%. A second functional site over 0.5 kb away was identified by deletion mutagenesis. These mutants also showed a greater than or equal to 96% loss of activity, indicating cooperativity between sites. The two classes of mutant protein displayed synergism of activity in a trans complementation assay. Effective actin microfilament bundle organization was also dependent on the combined function of both sites. Thus, fibroblast adhesion and intracellular response to the fibronectin cell-binding domain involve two synergistic sites, each of major quantitative importance. 相似文献
98.
In the previous experiment (Suzuki, H., Obara, M., Kuwayama, H., and Kanazawa, T. (1987) J. Biol. Chem. 262, 15448-15456), the Ca2+-ATPase of sarcoplasmic reticulum vesicles was labeled with N-iodoacetyl-N'-(5-sulfo-1-naphthyl)ethylenediamine without a loss of the catalytic activity. The main labeled site was Cys674. A large monophasic fluorescence drop occurred upon ATP binding to the catalytic site of the Ca2+-activated enzyme in the presence of K+. The present results show that this fluorescence drop is biphasic in the absence of K+. The first and rapid phase of this drop accounts for most of the fluorescence drop. This phase reflects a conformational change in the enzyme.ATP complex. The second and slow phase, being much smaller than the first phase, coincides with phosphoenzyme (EP) isomerization from the ADP-sensitive form to the ADP-insensitive form. This phase disappears when accumulation of ADP-insensitive EP is inhibited by K+ or when EP isomerization is prevented by the N-ethylmaleimide treatment. These results show that this phase reflects a conformational change upon EP isomerization. When free Ca2+ is chelated after EP formation from ATP, the fluorescence intensity is restored to the initial level without Ca2+. This restoration coincides with EP decomposition. This suggests that the fluorescence restoration reflects a conformational change upon hydrolysis of ADP-insensitive EP. This probability is supported by the concurrent occurrence of the Pi-induced fluorescence drop and EP formation from Pi. The results demonstrate that the fluorescence drop upon ATP binding is predominant in the fluorescence change throughout the catalytic cycle. 相似文献
99.
The effects of various peptides on the isolated pulmonary artery 总被引:2,自引:0,他引:2
Helical strips of pulmonary arteries from rabbits were tested for their responses to the following peptides: neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), substance P (SP), cholecystokinin-octapeptide (CCK), somatostatin (SS), bombesin, neurotensin and gastrin. SP, in the absence of active base-tension, and NPY both induced concentration-dependent contractions, while VIP and SP, in the presence of active base-tone, and CCK induced relaxation. The pD2 (-log ED50) was in the order of NPY greater than SP and SP greater than CCK greater than VIP. SS, bombesin, NT and gastrin had no effect. These findings suggest peptidergic involvement in the vessels. 相似文献
100.
M Taniguchi Y Tanigawa M Tsuchiya K Mishima S Obara K Yamada M Shimoyama 《Biochemical and biophysical research communications》1989,164(1):128-133
Arginine-specific ADP-ribosyltransferase from rabbit skeletal muscle sarcoplasmic reticulum was solubilized as the active form with trypsin. The enzyme was partially purified by subsequent chromatography, successively on DE-52, Con A-Sepharose and Sephadex G-75. An approximately 2,000-fold purification was achieved from the 105,000 x g supernatant of trypsin-treated membrane with a recovery of 2.8%. Dithiothreitol, which activates hen liver nuclear ADP-ribosyltransferase, inhibited the enzyme. 相似文献