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排序方式: 共有191条查询结果,搜索用时 31 毫秒
91.
92.
Shenping Wu Jun Liu Mary C. Reedy Richard T. Tregear Hanspeter Winkler Clara Franzini-Armstrong Hiroyuki Sasaki Carmen Lucaveche Yale E. Goldman Michael K. Reedy Kenneth A. Taylor 《PloS one》2010,5(9)
Background
Isometric muscle contraction, where force is generated without muscle shortening, is a molecular traffic jam in which the number of actin-attached motors is maximized and all states of motor action are trapped with consequently high heterogeneity. This heterogeneity is a major limitation to deciphering myosin conformational changes in situ.Methodology
We used multivariate data analysis to group repeat segments in electron tomograms of isometrically contracting insect flight muscle, mechanically monitored, rapidly frozen, freeze substituted, and thin sectioned. Improved resolution reveals the helical arrangement of F-actin subunits in the thin filament enabling an atomic model to be built into the thin filament density independent of the myosin. Actin-myosin attachments can now be assigned as weak or strong by their motor domain orientation relative to actin. Myosin attachments were quantified everywhere along the thin filament including troponin. Strong binding myosin attachments are found on only four F-actin subunits, the “target zone”, situated exactly midway between successive troponin complexes. They show an axial lever arm range of 77°/12.9 nm. The lever arm azimuthal range of strong binding attachments has a highly skewed, 127° range compared with X-ray crystallographic structures. Two types of weak actin attachments are described. One type, found exclusively in the target zone, appears to represent pre-working-stroke intermediates. The other, which contacts tropomyosin rather than actin, is positioned M-ward of the target zone, i.e. the position toward which thin filaments slide during shortening.Conclusion
We present a model for the weak to strong transition in the myosin ATPase cycle that incorporates azimuthal movements of the motor domain on actin. Stress/strain in the S2 domain may explain azimuthal lever arm changes in the strong binding attachments. The results support previous conclusions that the weak attachments preceding force generation are very different from strong binding attachments. 相似文献93.
K J Smith J D Wade A A Claasz L Otvos C Temelcos Y Kubota J M Hutson G W Tregear R A Bathgate 《Journal of peptide science》2001,7(9):495-501
The recently identified protein, insulin 3 (INSL3), has structural features that make it a bona fide member of the insulin superfamily. Its predicted amino acid sequence contains the classic two-peptide chain (A- and B-) structure with conserved cysteine residues that results in a disulphide bond disposition identical to that of insulin. Recently, the generation of insl3 knockout mice has demonstrated that testicular descent is blocked due to the failure of a specific ligament, the gubernaculum, to develop. The mechanism by which INSL3 exerts its action on the gubernaculum is currently unknown. The purpose of this study was to, for the first time, synthesize rat INSL3 and test its action on organ cultures of foetal rat gubernaculum. INSL3 also contains a cassette of residues Arg-X-X-X-Arg within the B-chain, a motif that is essential for characteristic activity of another related member of the superfamily, relaxin. Hence, the relaxin activity of rat INSL3 was also tested in two different relaxin bioassays. The primary structure of rat INSL3 was determined by deduction from its cDNA sequence and successfully prepared by solid phase peptide synthesis of the two constituent chains followed by their combination in solution. Following confirmation of its chemical integrity by a variety of analytical techniques, circular dichroism spectroscopy confirmed the presence of high beta-turn and alpha-helical content, with a remarkable spectral similarity to the synthetic ovine INSL3 peptide and to synthetic rat relaxin. The synthetic rat INSL3 bound with very low affinity to rat relaxin receptors and had no activity in a relaxin bioassay. Furthermore, it did not augment or antagonize relaxin activity. The rat INSL3 did however induce growth of foetal rat gubernaculum in whole organ cultures demonstrating that INSL3 has a direct action on this structure. 相似文献
94.
95.
96.
Jacobien J Hoogerwerf Gerritje JW van der Windt Dana C Blok Arie J Hoogendijk Alex F de Vos Cornelis van ‘t Veer Sandrine Florquin Koichi S Kobayashi Richard A Flavell Tom van der Poll 《Molecular medicine (Cambridge, Mass.)》2012,18(1):1067-1075
Pneumonia is a common cause of morbidity and mortality and the most frequent source of sepsis. Bacteria that try to invade normally sterile body sites are recognized by innate immune cells through pattern recognition receptors, among which toll-like receptors (TLRs) feature prominently. Interleukin-1 receptor (IL-1R)–associated kinase (IRAK)-M is a proximal inhibitor of TLR signaling expressed by epithelial cells and macrophages in the lung. To determine the role of IRAK-M in host defense against bacterial pneumonia, IRAK-M-deficient (IRAK-M−/−) and normal wild-type (WT) mice were infected intranasally with Klebsiella pneumoniae. IRAK-M mRNA was upregulated in lungs of WT mice with Klebsiella pneumonia, and the absence of IRAK-M resulted in a strongly improved host defense as reflected by reduced bacterial growth in the lungs, diminished dissemination to distant body sites, less peripheral tissue injury and better survival rates. Although IRAK-M−/− alveolar macrophages displayed enhanced responsiveness toward intact K. pneumoniae and Klebsiella lipopolysaccharide (LPS) in vitro, IRAK-M−/− mice did not show increased cytokine or chemokine levels in their lungs after infection in vivo. The extent of lung inflammation was increased in IRAK-M−/− mice shortly after K. pneumoniae infection, as determined by semiquantitative scoring of specific components of the inflammatory response in lung tissue slides. These data indicate that IRAK-M impairs host defense during pneumonia caused by a common gram-negative respiratory pathogen. 相似文献
97.
G R Sutherland E Baker V J Hyland D F Callen J A Close G W Tregear B A Evans R I Richards 《Cytogenetics and cell genetics》1988,48(4):205-207
The amino acid sequence of human prostate-specific antigen (APS) suggests that it is a member of the glandular kallikrein subfamily of serine proteases. In the mouse, the kallikrein-like family is localized in a single locus on chromosome 7, while other serine proteases are distributed over a variety of different chromosomes. To investigate the physical relationship between the human kallikrein genes, we have used in situ hybridization and Southern analysis of a human x mouse somatic cell hybrid panel to map the APS gene to 19q13, concordant with the renal kallikrein KLK1 gene. This finding indicates that APS is a member of a human kallikrein-like gene family with analogous organization to that of the mouse. 相似文献
98.
Denis Scanlon Jim Haralambidis Christina Southwell Janice Turton Geoffrey Tregear 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1984,336(1)
Synthetic oligodeoxyribonucleotides rangin from 11 to 37 nucleotides in length and with varying base compositions, prepared by both the phosphotriester and phosphite procedures, have been purified by ion-exchange high-performance liquid chromatography on Whatman Partisil 10/SAX columns using phosphate buffer gradients. The effects of different buffer systems on elution times and resolution have been evaluated. Oligomer composition and length had a marked effect on the resolution achieved. In general the use of formamide buffers gave the best results, particularly in the case of 2′-deoxyguanosine-rich sequences. These methods have also been successfully applied to the purification of mixtures of synthetic oligodeoxynucleotides. 相似文献
99.
The meridional spacing of the X-ray diffraction peak from the repeat of myosin along the thick filament of four muscles has been remeasured on the same apparatus. The frog sartorius gave a shorter repeat distance (143.7 A) than the three invertebrate muscles, which ranged from 144.9 to 145.4 A. These results confirm earlier measurements. Provided that the myosin molecules are staggered relative to one another by a constant 98 residues, it may be inferred that in vertebrate thick filaments part or all of the tail lies at a considerable angle to the filament axis, whereas in the invertebrates the angle is smaller. 相似文献
100.
A large set of 26 new reference transcriptomes dedicated to comparative population genomics in crops and wild relatives 下载免费PDF全文
Gautier Sarah Felix Homa Stéphanie Pointet Sandy Contreras François Sabot Benoit Nabholz Sylvain Santoni Laure Sauné Morgane Ardisson Nathalie Chantret Christopher Sauvage James Tregear Cyril Jourda David Pot Yves Vigouroux Hana Chair Nora Scarcelli Claire Billot Nabila Yahiaoui Roberto Bacilieri Bouchaib Khadari Michel Boccara Adéline Barnaud Jean‐Pierre Péros Jean‐Pierre Labouisse Jean‐Louis Pham Jacques David Sylvain Glémin Manuel Ruiz 《Molecular ecology resources》2017,17(3):565-580