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71.
72.
Most delta-endotoxins produced by Bacillus thuringiensis require proteolytic processing in order to become active. The in vitro and in vivo activation processes of Cry39A, a delta-endotoxin that is highly toxic to Anopheles stephensi, were investigated. Cry39A with a molecular mass of 72 kDa was processed in vitro into a 60 kDa fragment by trypsin and gut extract from A. stephensi larvae. N-terminal amino acid sequencing of the 60 kDa fragment revealed that trypsin and the protease(s) in the gut extract cleaved Cry39A between Arg(61) and Gly(62). In contrast, 40 and 25 kDa polypeptides were generated in vivo by intramolecular cleavage of the 60 kDa fragment in A. stephensi larvae. Further, a co-precipitation assay was used to investigate the binding property of the activated Cry39A to A. stephensi BBMV. Cry39A bound to A. stephensi BBMV specifically and did not compete with the Cry4Aa toxin. This indicated that the binding molecule(s) for Cry39A might differ from those for Cry4A. In addition, Cry39A preferentially bound to the Triton X-100-insoluble membrane fraction. 相似文献
73.
Hirano Y Hayashi H Iemura S Hendil KB Niwa S Kishimoto T Kasahara M Natsume T Tanaka K Murata S 《Molecular cell》2006,24(6):977-984
The 20S proteasome is a catalytic core of the 26S proteasome, a central enzyme in the degradation of ubiquitin-conjugated proteins. It is composed of 14 distinct gene products that form four stacked rings of seven subunits each, alpha(1-7)beta(1-7)beta(1-7)alpha(1-7). It is reported that the biogenesis of mammalian 20S proteasomes is assisted by proteasome-specific chaperones, named PAC1, PAC2, and hUmp1, but the details are still unknown. Here, we report the identification of a chaperone, designated PAC3, as a component of alpha rings. Although it can intrinsically bind directly to both alpha and beta subunits, PAC3 dissociates before the formation of half-proteasomes, a process coupled with the recruitment of beta subunits and hUmp1. Knockdown of PAC3 impaired alpha ring formation. Further, PAC1/2/3 triple knockdown resulted in the accumulation of disorganized half-proteasomes that are incompetent for dimerization. Our results describe a cooperative system of multiple chaperones involved in the correct assembly of mammalian 20S proteasomes. 相似文献
74.
Electromyographic analysis of walking in water in healthy humans 总被引:2,自引:0,他引:2
Masumoto K Takasugi S Hotta N Fujishima K Iwamoto Y 《Journal of PHYSIOLOGICAL ANTHROPOLOGY and Applied Human Science》2004,23(4):119-127
This study was designed to describe and clarify muscle activities which occur while walking in water. Surface electromyography (EMG) was used to evaluate muscle activities in six healthy subjects (mean age, 23.3 +/- 1.4 years) while they walked on a treadmill in water (with or without a water current) immersed to the level of the xiphoid process, and while they walked on a treadmill on dry land. The trials in water utilized the Flowmill which has a treadmill at the base of a water flume. Integrated EMG analysis was conducted for the quantification of muscle activities. In order to calculate the %MVC, the measurement of maximal voluntary contraction (MVC) of each muscle was made before the gait analysis, thus facilitating a comparison of muscle activities while walking in water with those on dry land. The %MVCs obtained from each of the tested muscles while walking in water, both with and without a water current, were all found to be lower than those obtained while walking on dry land at a level of heart rate response similar to that used when walking on dry land. Furthermore, the %MVCs while walking in water with a water current tended to be greater when compared to those while walking in water without a water current. In conclusion, the present study demonstrated that muscle activities while walking in water were significantly decreased when compared to muscle activities while walking on dry land, that muscle activities while walking in water tended to be greater with a water current than without, and that the magnitude of the muscle activity in water was relatively small in healthy humans. This information is important to design water-based exercise programs that can be safely applied for rehabilitative and recreational purposes. 相似文献
75.
76.
77.
Miho Iijima Kazuhiro Aiba Yoshimasa Tanaka Hideko Urushihara 《Journal of plant research》1998,111(1):93-96
Two cDNA fragments induced in developing zygotes ofDictyostelium discoideum were isolated by mRNA differential display. the relevant genes were also found to be expressed during asexual development,
suggesting that sexual and asexual development share common molecular mechanisms inD. discoideum. 相似文献
78.
Boris M. Hogema Jos C. Arents Rechien Bader Kevin Eijkemans Toshifumi Inada Hiroji Aiba & Pieter W. Postma 《Molecular microbiology》1998,28(4):755-765
The main mechanism causing catabolite repression by glucose and other carbon sources transported by the phosphotransferase system (PTS) in Escherichia coli involves dephosphorylation of enzyme IIAGlc as a result of transport and phosphorylation of PTS carbohydrates. Dephosphorylation of enzyme IIAGlc leads to 'inducer exclusion': inhibition of transport of a number of non-PTS carbon sources (e.g. lactose, glycerol), and reduced adenylate cyclase activity. In this paper, we show that the non-PTS carbon source glucose 6-phosphate can also cause inducer exclusion. Glucose 6-phosphate was shown to cause inhibition of transport of lactose and the non-metabolizable lactose analogue methyl-β- D -thiogalactoside (TMG). Inhibition was absent in mutants that lacked enzyme IIAGlc or were insensitive to inducer exclusion because enzyme IIAGlc could not bind to the lactose carrier. Furthermore, we showed that glucose 6-phosphate caused dephosphorylation of enzyme IIAGlc . In a mutant insensitive to enzyme IIAGlc -mediated inducer exclusion, catabolite repression by glucose 6-phosphate in lactose-induced cells was much weaker than that in the wild-type strain, showing that inducer exclusion is the most important mechanism contributing to catabolite repression in lactose-induced cells. We discuss an expanded model of enzyme IIAGlc -mediated catabolite repression which embodies repression by non- PTS carbon sources. 相似文献
79.
80.
Molecular cloning and nucleotide sequencing of the gene for E. coli cAMP receptor protein. 总被引:74,自引:17,他引:57 下载免费PDF全文
The crp gene of E. coli, which codes for cAMP receptor protein (CRP), has been cloned in the plasmid pBR322 on the basis of a genetic complementation. One of the recombinant plasmids, pHA1, was shown to direct the synthesis of CRP in a cell-free system. The location of the crp gene was determined by constructing subclones carrying various portions of pHA1. The nucleotide sequence of the crp gene has been determined. The coding region consists of 627 base pairs (bp), which specify a protein of 209 amino acids. The predicted amino acid sequence from the DNA sequence is consistent with the amino acid sequence partially known and the amino acid composition of CRP. After the coding region, there is a G-C rich inverted repeat sequence followed by a run of Ts, which could be a terminator of the crp gene. A possible promoter sequence was found about 180 bp upstream from the initiation codon and was shown to act as a promoter in vitro and in vivo. There are two dyad symmetry regions in a 167 bp leader sequence. 相似文献