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1.
Yongbo Hu Erin Webb Jasbir Singh Barry A Morgan James A Gainor Thomas D Gordon Teruna J Siahaan 《The Journal of biological chemistry》2002,277(10):8366-8371
The molecular basis of the substrate specificity of Clostridium histolyticum beta-collagenase was investigated using a combinatorial method. An immobilized positional peptide library, which contains 24,000 sequences, was constructed with a 7-hydroxycoumarin-4-propanoyl (Cop) fluorescent group attached at the N terminus of each sequence. This immobilized peptide library was incubated with C. histolyticum beta-collagenase, releasing fluorogenic fragments in the solution phase. The relative substrate specificity (k(cat)/K(m)) for each member of the library was determined by measuring fluorescence intensity in the solution phase. Edman sequencing was used to assign structure to subsites of active substrate mixtures. Collectively, the substrate preference for subsites (P(3)-P(4)') of C. histolyticum beta-collagenase was determined. The last position on the C-terminal side in which the identity of the amino acids affects the activity of the enzyme is P(4)', and an aromatic side chain is preferred in this position. The optimal P(1)'-P(3)' extended substrate sequence is P(1)'-Gly/Ala, P(2)'-Pro/Xaa, and P(3)'-Lys/Arg/Pro/Thr/Ser. The Cop group in either the P(2) or P(3) position is required for a high substrate activity with C. histolyticum beta-collagenase. S(2) and S(3) sites of the protease play a dominant role in fixing the substrate specificity. The immobilized peptide library proved to be a powerful approach for assessing the substrate specificity of C. histolyticum beta-collagenase, so it may be applied to the study of other proteases of interest. 相似文献
2.
In species in which boron (B) mobility is limited, B deficiency only occurs in growing plant organs. As a consequence of the highly localized patterns of plant growth and the general immobility of B it has been extremely difficult to determine the primary function of B in plants. In species in which B is phloem mobile, the removal of B from the growth medium results in the depletion of B present in mature leaves. Thus, it is possible to develop mature leaves with increasingly severe levels of B depletion, thereby overcoming the complications of experiments based on growing tissues. Utilizing this approach we demonstrate here that B depletion of mature plum (Prunus salicina) leaves did not result in any discernible change in leaf appearance, membrane integrity or photosynthetic capacity even though B concentrations were reduced to 6-8 µg/g dwt, which is less than 30% of the reported tissue B requirement. Boron depletion, however, results in a severe disruption of plant growth and metabolism in young growing tissues. This experimental evidence and theoretical considerations suggest that the primary and possibly sole function of B, is as a structural component of growing tissues. 相似文献
3.
Fish finders have already been widely available in the fishing market for a number of years.However,the sizes of these fishfinders are too big and their prices are expensive to suit for the research of robotic fish or mini-submarine.The goal of thisresearch is to propose a low-cost fish detector and classifier which suits for underwater robot or submarine as a proximity sensor.With some pre-condition in hardware and algorithms,the experimental results show that the proposed design has good per-formance,with a detection rate of 100 % and a classification rate of 94 %.Both the existing type of fish and the group behaviorcan be revealed by statistical interpretations such as hovering passion and sparse swimming mode. 相似文献
4.
Zenghui Hu Yingbai Shen Fanyi Shen Youqing Luo Xiaohua Su 《Trees - Structure and Function》2009,23(5):1003-1011
Interplant communication has been widely demonstrated in plants, especially in herbaceous plants. In this study, mechanical
damage was shown to affect the levels of pyrochatechol, chlorogenic acid, gallic acid and p-hydroxyl benzoic acid in poplar (Populus simonii × P. pyramidalis ‘Opera 8277’) cuttings, indicating the activation of defense response. In neighboring intact cuttings, the levels of those
phenolics also varied when compared to the control, suggesting the interplant communication between poplar cuttings. Three
volatiles, methyl jasmonate, methyl salicylate and benzothiazole, were detected in volatiles emitted from mechanically damaged
poplar cuttings. All of them can induce changes in the levels of four phenolics. Therefore, they could act as airborne signals
between P. simonii × P. pyramidalis ‘Opera 8277’ cuttings. The different change patterns of phenolic contents induced by different volatiles imply that the defense
response activated in neighboring plants may be regulated by multiple signaling pathways. The results also suggest that the
entire defense response of plants involves a variety of airborne signals in wound-induced volatiles. 相似文献
5.
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7.
P Hu G Parenti J Keulemans A T Hoogeveen 《Biochemical and biophysical research communications》1990,167(2):520-527
The catalytic and immunological properties of acid phosphatases (EC 3.1.3.2.) in different tissues were studied. It was demonstrated that high uptake forms of lysosomal enzymes like beta-galactosidase isolated from human platelets and bovine testis are mature enzymes, which have not lost their mannose-6phosphate marker. The results presented indicate that this phenomenon is related to a low activity or the complete absence of the lysosomal tartrate sensitive acid phosphatase activity in the tissues concerned. 相似文献
8.
9.
Purification and properties of formylglutamate amidohydrolase from Pseudomonas putida. 总被引:4,自引:3,他引:1 下载免费PDF全文
Formylglutamate amidohydrolase (FGase) catalyzes the terminal reaction in the five-step pathway for histidine utilization in Pseudomonas putida. By this action, N-formyl-L-glutamate (FG) is hydrolyzed to produce L-glutamate plus formate. Urocanate, the first product in the pathway, induced all five enzymes, but FG was able to induce FGase alone, although less efficiently than urocanate did. This induction by FG resulted in the formation of an FGase with electrophoretic mobility identical to that of the FGase induced by urocanate. A 9.6-kilobase-pair HindIII DNA fragment containing the P. putida FGase gene was cloned into the corresponding site on plasmid pBEU1 maintained in Escherichia coli. Insertion of the fragment in either orientation on the vector resulted in expression, but a higher level was noted in one direction, suggesting that the FGase gene can be expressed from either of two vector promoters with different efficiencies or from a single vector promoter in addition to a less efficient Pseudomonas promoter. FGase was purified 1,110-fold from the higher-expression clone in a yield of 10% through six steps. Divalent metal ions stimulated activity, and among those tested (Co, Fe, Zn, Ca, Ni, Cd, Mn, and Mg), Co(II) was the best activator, followed by Fe(II). FGase exhibited a Km of 14 mM for FG and a specific activity of 100 mumol/min per mg of protein in the presence of 5 mM substrate and 0.8 mM CoCl2 at 30 degrees C. The enzyme was maximally active in the range of pH 7 to 8. FGase was found to be a monomer of molecular weight 50,000. N-Acetyl-L-glutamate was not a substrate for the enzyme, but both it and N-formyl-L-aspartate were competitive inhibitors of formylglutamate hydrolysis, exhibiting Ki values of 6 and 9 mM, respectively. The absence of FGase activity as an integral part of histidine breakdown in most other organisms and the somewhat uncoordinated regulation of FGase synthesis with that of the other hut enzymes in Pseudomonas suggest that the gene encoding its synthesis may have evolved separately from the remaining hut genes. 相似文献
10.
E Hu K Watkins S Groshen S C Chen B Malloy R Agah P Nichols J Parker A Martin M G Boosalis 《Molecular biotherapy》1990,2(2):96-103
A phase I trial of interleukin-2 and interferon gamma combination treatment in patients with advanced malignancies was performed based on preclinical in vitro and in vivo data which demonstrated synergistic antitumor effect. The toxicities, immune parameters, and tumor responses are described. The clinical and biologic maximal tolerated doses were extrapolated from these data. 相似文献