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51.
We have previously shown that replacing the P1-site residue (Ala) of chicken ovomucoid domain 3 (OMCHI3) with a Met or Lys results in the acquisition of inhibitory activity toward chymotrypsin or trypsin, respectively. However, the inhibitory activities thus induced are not strong. In the present study, we introduced additional amino acid replacements around the reactive site to try to make the P1-site mutants more effective inhibitors of chymotrypsin or trypsin. The amino acid replacement Asp-->Tyr at the P2' site of OMCHI3(P1Met) resulted in conversion to a 35000-fold more effective inhibitor of chymotrypsin with an inhibitor constant (K(i)) of 1. 17x10(-11) M. The K(i) value of OMCHI3(P1Met, P2'Ala) indicated that the effect on the interaction with chymotrypsin of removing a negative charge from the P2' site was greater than that of introducing an aromatic ring. Similarly, enhanced inhibition of trypsin was observed when the Asp-->Tyr replacement was introduced into the P2' site of OMCHI3(P1Lys). Two additional replacements, Asp-->Ala at the P4 site and Arg-->Ala at the P3' site, made the mutant a more effective inhibitor of trypsin with a K(i) value of 1. 44x10(-9) M. By contrast, Arg-->Ala replacement at the P3' site of OMCHI3(P1Met, P2'Tyr) resulted in a greatly reduced inhibition of chymotrypsin, and Asp-->Ala replacement at the P4 site produced only a small change when compared with a natural variant of OMCHI3. These results clearly indicate that not only the P1-site residue but also the characteristics, particularly the electrostatic properties, of the amino acid residues around the reactive site of the protease inhibitor determine the strength of its interactions with proteases. Furthermore, amino acids with different characteristics are required around the reactive site for strong inhibition of chymotrypsin and trypsin. 相似文献
52.
Z Zhong A Toukdarian D Helinski V Knauf S Sykes J E Wilkinson C O'Bryne T Shea C DeLoughery R Caspi 《Applied and environmental microbiology》2001,67(12):5771-5779
An agar-degrading marine bacterium identified as a Microscilla species was isolated from coastal California marine sediment. This organism harbored a single 101-kb circular DNA plasmid designated pSD15. The complete nucleotide sequence of pSD15 was obtained, and sequence analysis indicated a number of genes putatively encoding a variety of enzymes involved in polysaccharide utilization. The most striking feature was the occurrence of five putative agarase genes. Loss of the plasmid, which occurred at a surprisingly high frequency, was associated with loss of agarase activity, supporting the sequence analysis results. 相似文献
53.
Y. Salas A. Márquez J. Canelón Y. Perazzo V. Colmenárez J. A. López 《Mycopathologia》2012,174(5-6):511-517
Pythium insidiosum is a pathogenic oomycete known since 1890 that causes pythiosis in mammals. In this report, seven P. insidiosum isolates were recovered from Venezuelan horses and were characterized. The strains were recovered from biopsied tissues and kunkers collected from granulomatous masses located on the hind limb and from a nodular lesion in the left upper eyelid, which decrease the ability of the horses to be used for working purposes. The methods used to identify P. insidiosum isolates were based on the production of sporangia and zoospores, histopathology and PCR assay. To further characterize these strains, portions of the 18S rRNA genes of the seven isolates were sequenced. The sequences showed high homology to previously described P. insidiosum DNA sequences available in GenBank. Similar studies based on the morphological, histological and molecular data identified the etiological agent in samples of granulomatous lesions in these equines as P. insidiosum. In America, the infection has been diagnosed more frequently in equines of Brazil, Colombia, Costa Rica and the United States of America. 相似文献
54.
J R Silkensen A P Skubitz K M Skubitz M E Rosenberg 《The journal of peptide research》1999,54(5):449-457
Expression of the glycoprotein clusterin is markedly increased following tissue injury. One function of clusterin is to promote cell interactions which are perturbed in these pathologic settings. Clusterin causes cell aggregation and adhesion in vitro yet the molecular mechanism for this effect is not known. In order to identify the active site(s) of clusterin, 34 peptides, each 15 amino acid residues in length, were synthesized from hydrophilic regions of human clusterin. When studied individually, none of the peptides caused aggregation of LLC-PK1 cells, a porcine renal epithelial cell line. However, two out of the 34 peptides inhibited clusterin-induced cell aggregation in a dose-dependent manner. Scrambled versions of these two 'active' peptides did not inhibit cell aggregation. Seven peptides promoted cell adhesion. In conclusion, these findings provide evidence for novel amino acid sequences mediating clusterin-induced renal cell interactions. 相似文献
55.
Wendy A. Douglass Robert H. Hyland Christopher D. Buckley Aymen Al-Shamkhani Jacqueline M. Shaw Sarah L. Scarth David L. Simmons S.K.Alex Law 《FEBS letters》1998,440(3):125
The cysteine-rich region (CRR) of the β2 integrin subunit was replaced by that of β1 to give the chimera β2NV1. β2NV1 can combine with αL to form a variant leukocyte-function-associated antigen (LFA)-1 on COS cell surface, suggesting that the specificity of the β2 interaction with αL does not lie in the CRR. Unlike those expressing wild-type LFA-1, COS cells expressing αLβ2NV1 are constitutively active in intercellular adhesion molecule (ICAM)-1 adhesion. These results suggest that activation of LFA-1 involves the release of an intramolecular constraint, which is maintained, in part, by the authentic β2 CRR. 相似文献
56.
Self-consistent steady-state axisymmetric configurations of a plasma envelope with a uniform anisotropic conductivity around a rotating magnetized spherical body are considered. A set of electrodynamic and magnetohydrodynamic equations is analyzed under the assumption that the mass velocity of a moving weakly ionized plasma has only the azimuthal component. The equations describing the profile of the angular frequency of the rotating plasma envelope, the magnetic field, the conduction currents, and the plasma density distribution are solved in the limit of a strong anisotropy of the conductivity of a weakly ionized gas. The applicability of the results obtained to a qualitative interpretation of the phenomena occurring in the plasmaspheres of magnetized planets is discussed. 相似文献
57.
I. N. Semenchuk L. A. Taranova A. A. Kalenyuk P. V. Il’yasov A. N. Reshetilov 《Applied Biochemistry and Microbiology》2000,36(1):69-72
The operating and storage stability of a receptor element of an amperometric biosensor based on thePseudomonas rathonis strain T capable of degrading surfactants was tested. Microbial cells were immobilized by incorporation in gels (agar, agarose,
and calcium-alginate), polyvinyl alcohol membrane, adhesion to Chromatographic paper GF/A, or by cross-linking induced by
glutaric aldehyde. Incorporation of microbial cells in agar gel provides long-standing conservation of their activity and
viability during measurements of high concentrations of surfactants and allows the receptor element of the biosensor to be
rapidly recovered after measurements. 相似文献
58.
D S Faffe G H Silva P M Kurtz E M Negri V L Capelozzi P R Rocco W A Zin 《Journal of applied physiology》2001,90(4):1400-1406
The dynamic mechanical properties of lung tissue and its contents of collagen and elastic fibers were studied in strips prepared from mice instilled intratracheally with saline (C) or silica [15 (S15) and 30 days (S30) after instillation]. Resistance, elastance, and hysteresivity were studied during oscillations at different frequencies on S15 and S30. Elastance increased from C to silica groups but was similar between S15 and S30. Resistance was augmented from C to S15 and S30 and was greater in S30 than in S15 at higher frequencies. Hysteresivity was higher in S30 than in C and S15. Silica groups presented a greater amount of collagen than did C. Elastic fiber content increased progressively along time. This increment was related to the higher amount of oxytalan fibers at 15 and 30 days, whereas elaunin and fully developed elastic fibers were augmented only at 30 days. Silicosis led not only to pulmonary fibrosis but also to fibroelastosis, thus assigning a major role to the elastic system in the silicotic lung. 相似文献
59.
V. Grant 《Plant Systematics and Evolution》2001,230(1-2):89-96
This paper deals with the use of cladistic methods and cladograms in phylogeny reconstruction in plant groups containing
numerous taxa. How accurate are the cladograms as to details? Accuracy tests at the level of details require an independently
known phylogeny, which excludes most plant groups, but such tests can be carried out in domesticated and experimental plant
groups which have documented pedigrees. Four such tests are known and are presented here: a new case in Gilia and three previously published cases in Avena, Hordeum, and Helianthus. The four cases include domesticated and experimental plants, use of morphological and molecular evidence, and presence of
dichotomous as well as reticulate phylogenies. The cladograms of the four plant groups all differ in significant details from
the known pedigrees. These results are discussed in relation to problems of interpretation of cladograms.
Received March 21, 2000 Accepted August 16, 2001 相似文献
60.
S. I. Tkachenko A. R. Mingaleev V. M. Romanova A. E. Ter-Oganes’yan T. A. Shelkovenko S. A. Pikuz 《Plasma Physics Reports》2009,35(9):734-753
Distribution of matter in the discharge channel formed upon a nanosecond electrical explosion of a single wire in air and
vacuum was studied experimentally. Simultaneous use of optical, UV, and X-ray diagnostics made it possible to distinguish
qualitatively different regions of the discharge channel, such as the current-carrying layers and the region occupied by a
weakly conducting cold plasma. Several series of experiments with 25-μm-diameter 12-mm-long wires made of different materials
were performed. The charging voltage and the current amplitude were varied in the ranges of U
0 = 10–20 kV and I
max ∼ 5–10 kA, respectively. Explosion regimes with a current pause and with and without current interruption, as well as with
wire preheating in air and vacuum, were studied. Shadow and schlieren images of the discharge channel were obtained using
optical probing at the second harmonic of a YAG: Nd+3 laser (λ = 0.532 μm, τ ∼ 10 ns). In the experiments carried out in vacuum, X-ray images of the discharge channel were also
obtained using an X-pinch as a point source of probing radiation and UV images were recorded using a four-frame MCP camera. 相似文献