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1.
The zymogen alpha-chymotrypsinogen A is bound to mesoporous silica MCM-41 with a protein loading of 170 mg/g solid (MCM-Z) by a simple stirring in aqueous tris-HCl buffer (pH 7.2). The bound zymogen is then activated with trypsin to obtain alpha-chymotrypsin immobilized on MCM-41 (MCM-E.I) that displays an effective enzyme activity corresponding to 65 mg protein/g of solid support (3250 BTEE units/g). A direct immobilization of commercially available alpha-chymotrypsin (MCM-E.II) gives lower loading (1250 BTEE units/g). Protein content of the solid support after immobilization is confirmed by thermogravimetric analysis (TGA). The enzyme is tightly bound to the support and can be used over 100 recycles over 1 week in aqueous as well as reverse micellar media. The immobilized enzyme (MCM-E.I) has been used for resolution of N-acetyl-dl-amino acid esters and racemic trans-4-methoxy-3-phenylglycidic acid (PGA) methyl ester. 相似文献
2.
Gary S. Ma Inmaculada Lopez-Sanchez Nicolas Aznar Nicholas Kalogriopoulos Shabnam Pedram Krishna Midde Theodore P. Ciaraldi Robert R. Henry Pradipta Ghosh 《Molecular biology of the cell》2015,26(23):4209-4223
Insulin resistance (IR) is a metabolic disorder characterized by impaired insulin signaling and cellular glucose uptake. The current paradigm for insulin signaling centers upon the insulin receptor (InsR) and its substrate IRS1; the latter is believed to be the sole conduit for postreceptor signaling. Here we challenge that paradigm and show that GIV/Girdin, a guanidine exchange factor (GEF) for the trimeric G protein Gαi, is another major hierarchical conduit for the metabolic insulin response. By virtue of its ability to directly bind InsR, IRS1, and phosphoinositide 3-kinase, GIV serves as a key hub in the immediate postreceptor level, which coordinately enhances the metabolic insulin response and glucose uptake in myotubes via its GEF function. Site-directed mutagenesis or phosphoinhibition of GIV-GEF by the fatty acid/protein kinase C-theta pathway triggers IR. Insulin sensitizers reverse phosphoinhibition of GIV and reinstate insulin sensitivity. We also provide evidence for such reversible regulation of GIV-GEF in skeletal muscles from patients with IR. Thus GIV is an essential upstream component that couples InsR to G-protein signaling to enhance the metabolic insulin response, and impairment of such coupling triggers IR. We also provide evidence that GIV-GEF serves as therapeutic target for exogenous manipulation of physiological insulin response and reversal of IR in skeletal muscles. 相似文献
3.
Sekhar PN Kishor PB Reddy LA Mondal P Dash AK Kar M Mohanty S Sabat SC 《In silico biology》2006,6(5):435-447
Homology modeling of the catalase, CatC cloned and sequenced from rice (Oryza sativa L., cv Ratna an Indica cultivar) has been performed based on the crystal structure of the catalase CatF (PDB code 1m7s) by using the software MODELLER. With the aid of molecular mechanics and molecular dynamics methods, the final model is obtained and is further assessed by PROCHECK and VERIFY - 3D graph, which show that the final refined model is reliable. With this model, a flexible docking study with the hydrogen peroxide, the substrate for catalase, is performed and the results indicate that Arg310, Asp343 and Arg346 in catalase are three important determinant residues in binding as they have strong hydrogen bonding contacts with the substrate. These hydrogen-bonding interactions play an important role for the stability of the complex. Our results may be helpful for further experimental investigations. 相似文献
4.
Catalase plays a central role in plant stress responses but is highly susceptible to photoinhibition. A rice catalase-B protein
avoiding photoinhibition was developed by mutagenesis of specific amino acids: Leu-189 to Trp-189 and His-225 to Thr-225 and
then recombinantly expressed in E. coli. In addition, the site specific mutation also induced 2–2.5-fold increase in enzyme velocity with high affinity for its substrate
and showed nearly a 3-fold lower K
m
than the wild protein. These characteristic of mutated rice catalase-B is highly promising in transgenic research to increase
plant productivity under stress conditions. 相似文献
5.
6.
A. Murugadoss Manoranjan Kar Renu Pasricha Arun Chattopadhyay 《Plasmonics (Norwell, Mass.)》2009,4(2):161-170
In this paper, we report the spontaneous formation of fibrous structures consisting of assemblies of Au–Ag core-shell nanoparticles
(NPs) from a solution consisting of Au–Ag core-shell NPs and l-ascorbic acid (AA). AA acted both as the reducing agent for the generation of NPs and also as the mediator for the formation
of fibers. The process of fiber formation involved three steps—reduction of HAuCl4 to Au NPs by AA, subsequent formation of Au–Ag core-shell NPs after addition of AgNO3, and spontaneous formation of fibers from the mixtures in water. It took typically about 30 days to form complete fibers
that are of lengths of several hundred micrometers to millimeters, although nanofibers started forming from the first day
of solution preparation. The width of each of these fibers was typically about 1–4 μm with length of each segment of fiber
bundle, on the order of 40 μm. Formation of fibers was also observed in absence of AgNO3. These fibers consisted of Au NPs and polymer of AA degradation products and were not electrically conducting. Also, low
concentrations of AgNO3 produced fibers with low electrical conductivity. However, it was observed that increase in the amount of AgNO3 leads to the formation of fibers that were electrically conducting with conductivity values in the range of metallic conductivity.
Spectroscopic and electron microscopic investigations were carried out to establish the formation of fibers. The details of
fiber formation mechanism under different conditions and electrical conductivities of the fibers are discussed in the article. 相似文献
7.
Krishna Midde Ryan Rich Ashwini Saxena Ignacy Gryczynski Julian Borejdo Hriday K. Das 《Cell biochemistry and biophysics》2014,70(2):923-932
Presenilin-1 (PS1) protein acts as passive ER Ca2+ leak channels that facilitate passive Ca2+ leak across ER membrane. Mutations in the gene encoding PS1 protein cause neurodegeneration in the brains of patients with familial Alzheimer’s disease (FAD). FADPS1 mutations abrogate the function of ER Ca2+ leak channel activity in human neuroblastoma SK-N-SH cells in vitro (Das et al., J Neurochem 122(3):487–500, 2012) and in mouse embryonic fibroblasts. Consequently, genetic deletion or mutations of the PS1 gene cause calcium (Ca2+) signaling abnormalities leading to neurodegeneration in FAD patients. By analogy with other known ion channels it has been proposed that the functional PS1 channels in ER may be multimers of several PS1 subunits. To test this hypothesis, we conjugated the human PS1 protein with an NH2-terminal YFP-tag and a COOH-terminal CFP-tag. As expected YFP–PS1, and PS1–CFP were found to be expressed on the plasma membranes by TIRF microscopy, and both these fusion proteins increased ER Ca2+ leak channel activity similar to PS1 (WT) in SK-N-SH cells, as determined by functional calcium imaging. PS1–CFP was either expressed alone or together with YFP–PS1 into SK-N-SH cell line and the interaction between YFP–PS1 and PS1–CFP was determined by Förster resonance energy transfer analysis. Our results suggest interaction between YFP–PS1 and PS1–CFP confirming the presence of a dimeric or multimeric form of PS1 in SK-N-SH cells. Lateral diffusion of PS1–CFP and YFP–PS1 in the plasma membrane of SK-N-SH cells was measured in the absence or in the presence of glycerol by fluorescence correlation spectroscopy to show that both COOH-terminal and NH2-terminal of human PS1 are located on the cytoplasmic side of the plasma membrane. Therefore, we conclude that both COOH-terminal and NH2-terminal of human PS1 may also be oriented on the cytosolic side of ER membrane. 相似文献
8.
A unique Mycobacterium ESX-1 protein co-secretes with CFP-10/ESAT-6 and is necessary for inhibiting phagosome maturation 总被引:3,自引:0,他引:3
Xu J Laine O Masciocchi M Manoranjan J Smith J Du SJ Edwards N Zhu X Fenselau C Gao LY 《Molecular microbiology》2007,66(3):787-800
The ESX-1 secretion system plays a critical role in the virulence of Mycobacterium tuberculosis and M. marinum. To date, three proteins are known to be secreted by ESX-1 and necessary for virulence, two of which are CFP-10 and ESAT-6. The ESX-1 secretion and the virulence mechanisms are not well understood. In this study, we have examined the M. marinum secretomes and identified four proteins specific to ESX-1. Two of those are CFP-10 and ESAT-6, and the other two are novel: MM1553 (homologous to Rv3483c) and Mh3881c (homologous to Rv3881c). We have shown that Mh3881c, CFP-10 and ESAT-6 are co-dependent for secretion. Mh3881c is being cleaved at close to the C-terminus during secretion, and the C-terminal portion is critical to the co-dependent secretion, the ESAT-6 cellular levels, and interaction with ESAT-6. The co-dependent secretion is required for M. marinum intracellular growth in macrophages, where the Mh3881c C-terminal portion plays a critical role. The role of the co-dependent secretion in intracellular growth correlates with its role in inhibiting phagosome maturation. Both the secretion and the virulence defects of the Mh3881c mutant are complemented by Mh3881c or its M. tuberculosis homologue Rv3881c, suggesting that in M. tuberculosis, Rv3881c has similar functions. 相似文献
9.
Genotyping of Novel SNPs in BMPR1B,BMP15, and GDF9 Genes for Association with Prolificacy in Seven Indian Goat Breeds 总被引:1,自引:0,他引:1
Sonika Ahlawat Rekha Sharma Manoranjan Roy Sanjay Mandakmale Ved Prakash M S Tantia 《Animal biotechnology》2016,27(3):199-207
Goats form the backbone of rural livelihood and financial security systems in India but their population is showing decreasing trend. Improvement of reproductive traits such as prolificacy offers a solution to stabilize the decreasing goat population and to meet the nutritional needs of growing human population. In the present study, six novel SNPs in three candidate genes for prolificacy (BMPR1B, BMP15, and GDF9) were genotyped in seven breeds of Indian goats to evaluate their association with litter size. Tetra primer ARMS-PCR and PCR-RFLP based protocols were developed for genotyping six novel SNPs, namely, T(-242)C in BMPR1B; G735A and C808G in BMP15; and C818T, A959C, and G1189A in GDF9 gene. The effect of breed was highly significant (p ≤ 0.01) on litter size but the effect of genotype was nonsignificant. The effect of parity on litter size was also significant in the prolific Black Bengal breed. The litter size differences observed between breeds are attributed to breed differences. Novel mutations observed at different loci in GDF9, BMP15, and BMPR1B genes do not contribute to the reproductive capability of the investigated breeds. Further studies with more number of breeds and animals exploring association of these novel SNPs with reproductive traits may be fruitful. 相似文献
10.
The Belousov-Zhabotinskii reaction has been modelled by Field and Noyes [5] as a pair of nonlinear parabolic equations. Previous studies of these, both theoretical and numerical, have assumed wave solutions travelling with constant velocity leading to a simplification of the mathematical model in the form of a system of ordinary differential equations. In the present study a finite element Galerkin method is used directly on the original parabolic system for a range of parameter values. 相似文献