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1.
Springs SL  Bass SE  McLendon GL 《Biochemistry》2000,39(20):6075-6082
A general understanding of how cytochromes evolve within a fixed structure to optimize redox potential for specific bioenergetic processes does not exist. Toward this end, a library approach is used to investigate the range and distribution of redox potential which occurs when all sequence space available through mutation at two positions is examined within a fixed structural motif. Random mutation of Phe61 and Phe65 of cytochrome b562 (E. coli), and subsequent examination of a statistically significant sampling of this library, demonstrates that the redox potential can vary over 100 mV (>25% of the known accessible potential in native proteins with axial His-Met ligation) through mutation at these two positions. The redox potential of the wild-type protein occurs at an extremum of the distribution observed, indicating that Phe61 and Phe65 were most likely naturally selected to differentially stabilize the reduced state of the protein. At the other extremum, a compositionally conservative set of mutations (F61I, F65Y) leads to a 100 mV shift in the redox equilibrium toward the oxidized state. NMR analyses indicate that a charge-dipole interaction which results from mutation of phenylalanine to tyrosine at position 65 may be responsible.  相似文献   
2.
A novel approach is described for generating reactive oxidizing centers in heme proteins, with zinc hemoglobin (Zn Hb) and zinc cytochrome c (Zn cyt c) used as examples. The reaction of 3Zn* Hb with [CoIII(NH3)5 Cl]2+, and of 3Zn* cyt c with methyl viologen are described. In the case of Zn Hb the cation radical produced decays with a rate constant of k3 = 2400s-1. Using this value the rate of the reaction (formula; see text) can be calculated to be 4500s-1.  相似文献   
3.
WhileEscherichia coli is common as a commensal organism in the distal ileum and colon, the presence of colonization factors (CF) on pathogenic strains ofE. coli facilitates attachment of the organism to intestinal receptor molecules in a species- and tissue-specific fashion. After the initial adherence, colonization occurs, and the involvement of additional virulence determinants leads to illness. EnterotoxigenicE. coli (ETEC) is the most extensively studied of the five categories ofE. coli that cause diarrheal disease, and has the greatest impact on health worldwide. ETEC can be isolated from domestic animals and humans. The biochemistry, genetics, epidemiology, antigenic characteristics, and cell and receptor binding properties of ETEC have been extensively described. Another major category, enteropathogenicE. coli (EPEC), has virulence mechanisms, primarily effacement and cytoskeletal rearrangement of intestinal brush borders, that are distinct from ETEC. An EPEC CF receptor has been purified and characterized as a sialidated transmembrane glycoprotein complex directly attached to actin, thereby associating CF-binding with host-cell response. Three, additional categories ofE. coli diarrheal disease, their colonization factors and their host cell receptors are discussed. It appears that biofilms exist in the intestine in a manner similar to oral bacterial biofilms, and thatE. coli is part of these biofilms as both commensals and pathogens.Abbreviations CF colonization factor - CFA Colonization Factor Antigen - CS coli-surface-associated antigen - EAggEC enteroaggregativeE. coli - ECDD E. coli diarrheal disease - EHEC enterohemorrhagicE. coli - EIEC enteroinvasiveE. coli - EPEC enteropathogenicE. coli - ETEC enterotoxigenicE. coli - Gal galactose - GalNAc N-acetyl galactosamine - LT heat-labile toxin - NeuAc N-acetyl neuraminic acid - PCF Putative colonization factor - RBC red blood cells - SLT Shiga-like toxin - ST heat-stable toxin  相似文献   
4.
The relationship between soil nitrogen (N) availability and plant community structure was investigated in old-fields in the shortgrass steppe of Colorado. Nitrogen availability was manipulated by N or sucrose additions for 4 years at three old-fields (early-seral, mid-seral, and late-seral) and at an uncultivated control site. The addition of N generally resulted in increased abundance of annual forbs and grasses relative to perennials at all of the previously cultivated sites. Conversely, experimental reduction of N availability generally increased the relative abundance of perennials. Despite a lack of detectable differences in N mineralization between sites and treatments, ion-exchange resin bags confirmed that sucrose additions reduced plant-available N and that N additions increased plant-available N. This was evidenced further by similar observations for plant tissue N content. The degree to which N additions increased N availability at the various sites supported the idea that late-seral plant communities are less effective at N capture relative to earlier-seral communities. The mid-seral old-field had the lowest rates of litter decomposition and a relatively large accumulation of litter on the soil surface. This mid-seral old-field was dominated by an exotic annual grass (Bromus tectorum), which appears to be a major hindrance to redevelopment of the plant-soil system. By experimentally reducing N availability at this stage, we were able, in 4 years, to change the plant community into one that more closely resembled the late-seral community. We also observed that the natural recruitment of weedy annual species on the uncultivated site during an unusually wet year was suppressed by reducing N availability. Our results suggest that available N is an important factor controlling the rate and course of plant and soil community redevelopment on abandoned croplands in the shortgrass steppe, and that manipulation of N availability might be useful in restoration of rangeland vegetation. Received 19 May 1998; accepted 27 August 1999.  相似文献   
5.
Fluorescent lanthanide ions, with large Stokes shifts and narrow emission bands, are excellent tools for the development of FRET-based assays. In this work, a terbium ion is tethered to a peptide which binds to the BIR3 domain of XIAP, an anti-apoptotic protein. Excitation of tryptophan and tyrosine residues in the BIR3 domain causes the peptide bound terbium ion to fluoresce relative to its distance from these aromatic residues. By developing ligands with terbium ions tethered at different residues, the relative terbium emission can be used to "map" the aromatic residues within the ligand binding pocket.  相似文献   
6.
    
The field of biomarkers is a growing one, particularly in osteoarthritis (OA). OA is the most common disabling condition in older persons and a major cause of morbidity. While the debate continues about which of the involved tissues - cartilage, bone or synovium - is the most important in OA aetiology, there is no doubt that the three develop abnormalities in concert; perhaps a truly useful biomarker will reflect just that. While efforts continue to identify reliable biomarkers useful for characterising the status, prognosis and measurement of treatment response in OA, combining existing biomarkers to improve their accuracy looks promising.  相似文献   
7.
8.
  总被引:6,自引:0,他引:6  
Studies of classic zinc-finger peptides over the past 15 years have offered insights into the coupled processes of metal binding and protein folding. Within the past two years, this insight has been used to increase our understanding of the importance of first and second shell contributions (i.e. contributions from direct and indirect metal ligands) to metal binding and protein-folding stability, and led to advances in de novo protein design and protein redesign.  相似文献   
9.
J Feitelson  G McLendon 《Biochemistry》1991,30(20):5051-5055
It has previously been shown that the rates and activation energies for migration molecules of different sizes through myoglobin are very similar. The results were interpreted in terms of conformational changes in the protein structure that facilitate the passage of the different molecules to a similar extent. Here we ask whether the quaternary structural changes that accompany the binding of ligands (O2 or CO) to hemoglobin might influence the migration rate from the solution into the protein's binding site. As a model for the R state of hemoglobin, we used the protein in which the Fe protoporphyrin (FePP) in the alpha subunit was substituted by Zn protoporphyrin (ZnPP) and the oxidized heme was ligated by CN-. The T state of hemoglobin was represented by the protein in which all four FePP groups were substituted by ZnPP. The quenching rate of the excited ZnPP triplet state within the hemoglobin by oxygen, methyl viologen, and anthraquinonesulfonate served as a measure of the migration rate through the protein into the binding site. It was found that the activation energies for all three quenchers were very similar and closely resembled those in myoglobin, suggesting that the migration rates are determined by the subunit structure only and that the quaternary configurational changes do not influence the quenching rates. The implications of the results for electron transfer in proteins are briefly discussed.  相似文献   
10.
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