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81.
Shin NY Dise RS Schneider-Mergener J Ritchie MD Kilkenny DM Hanks SK 《The Journal of biological chemistry》2004,279(37):38331-38337
Crk-associated substrate (p130(CAS) or CAS) is a major integrin-associated Src substrate that undergoes tyrosine phosphorylation at multiple YXXP motifs in its substrate domain (SD) to create docking sites for SH2-containing signaling effectors. Notably, recruitment of Crk adaptor proteins to the CAS SD sites is implicated in promoting cell migration. However, it is unclear which or how many of the 15 CAS SD YXXP tyrosines are critically involved. To gain a better understanding of CAS SD function, we assessed the signaling capacity of individual YXXP motifs. Using site-directed mutagenesis combined with tryptic phosphopeptide mapping, we determined that the ten tyrosines in YXXP motifs 6-15 are the major sites of CAS SD phosphorylation by Src. Phosphopeptide binding assays showed that all of these sites are capable of binding the Crk SH2 domain. To evaluate the requirement for CAS YXXP sites in stimulating cell migration, a series of phenylalanine substitution variants were expressed in CAS -/- mouse embryo fibroblasts. CAS expression enhanced the rate of cell migration into a monolayer wound in a manner dependent on the major sites of Src phosphorylation. Effective wound healing was achieved by CAS variants containing as few as four of the major sites, indicating sufficiency of partial SD signaling function in this cell migration response. 相似文献
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Crk-associated substrate tyrosine phosphorylation sites are critical for invasion and metastasis of SRC-transformed cells 总被引:6,自引:0,他引:6
Brábek J Constancio SS Siesser PF Shin NY Pozzi A Hanks SK 《Molecular cancer research : MCR》2005,3(6):307-315
Crk-associated substrate (CAS, p130Cas) is a major tyrosine phosphorylated protein in cells transformed by v-crk and v-src oncogenes. We recently reported that reexpression of CAS in CAS-deficient mouse embryo fibroblasts transformed by oncogenic Src promoted an invasive phenotype associated with enhanced cell migration through Matrigel, organization of actin into large podosome ring and belt structures, activation of matrix metalloproteinase-2, and elevated tyrosine phosphorylation of the focal adhesion proteins FAK and paxillin. We have now extended these studies to examine the mechanism by which CAS achieves these changes and to evaluate the potential role for CAS in promoting in vivo tumor growth and metastasis. Whereas the presence or absence of CAS did not alter the primary growth of subcutaneous-injected Src-transformed mouse embryo fibroblasts, CAS expression was required to promote lung metastasis following removal of the primary tumor. The substrate domain YxxP tyrosines, the major sites of CAS phosphorylation by Src that mediate interactions with Crk, were found to be critical for promoting both invasive and metastatic properties of the cells. The ability of CAS to promote Matrigel invasion, formation of large podosome structures, and tyrosine phosphorylation of Src substrates, including FAK, paxillin, and cortactin, was also strictly dependent on the YxxP tyrosines. In contrast, matrix metalloproteinase-2 activation was most dependent on the CAS SH3 domain, whereas the substrate domain YxxP sites also contributed to this property. Thus multiple CAS-mediated signaling events are implicated in promoting invasive and metastatic properties of Src-transformed cells. 相似文献
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Sexual dimorphism in insect antennal structure is often attributed to differences between the sexes in sensitivity to pheromones, the antennae of one sex being more elaborately structured (for example, plumose). Males of the family Cerambycidae (order Coleoptera) often have longer antennae than females, but of a similar general structure, suggesting that selective factors other than sensitivity to pheromones are at work. Both sexes of the eucalyptus longhorned borer, a cerambycid, were attracted to eucalyptus logs that were larval hosts. There, males located females by antennal contact, and male mating success therefore depended on the walking rate and width of the antennal spread. Elongate antennae may benefit males by increasing antennal spread width, but have no such advantage for females, suggesting an evolutionary explanation for sexual dimorphism. 相似文献
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Emerson S. Lacey Jardel A. Moreira Jocelyn G. Millar Ann M. Ray & Lawrence M. Hanks 《Entomologia Experimentalis et Applicata》2007,122(2):171-179
Adult male Neoclytus mucronatus mucronatus (F.) (Coleoptera: Cerambycidae: Cerambycinae) were observed to display behaviors identical to calling behaviors of the congener N. acuminatus acuminatus F., males of which produce an aggregation pheromone. Odors collected from male N. m. mucronatus contained one major male‐specific compound, identified as (R)‐3‐hydroxyhexan‐2‐one. Bioassays determined that both sexes were weakly attracted to racemic 3‐hydroxy‐2‐hexanone. Further field trials determined that enantiomerically enriched (R)‐3‐hydroxyhexan‐2‐one (94% ee) attracted more beetles of both sexes than did the racemic blend. This aggregation pheromone is produced by glands that discharge through pores lying within shallow cuticular depressions in the pronotum of male N. m. mucronatus. 相似文献
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J H Hanks 《Microbiology and molecular biology reviews》1966,30(1):114-135
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