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Unique Interactions (unique + interaction)
Selected AbstractsUnique interactions between the chromophore and glutamate 16 lead to far-red emission in a red fluorescent proteinPROTEIN SCIENCE, Issue 2 2009Xiaokun Shu Abstract mPlum is a far-red fluorescent protein with emission maximum at ,650 nm and was derived by directed evolution from DsRed. Two residues near the chromophore, Glu16 and Ile65, were previously revealed to be indispensable for the far-red emission. Ultrafast time-resolved fluorescence emission studies revealed a time dependent shift in the emission maximum, initially about 625 nm, to about 650 nm over a period of 500 ps. This observation was attributed to rapid reorganization of the residues solvating the chromophore within mPlum. Here, the crystal structure of mPlum is described and compared with those of two blue shifted mutants mPlum-E16Q and -I65L. The results suggest that both the identity and precise orientation of residue 16, which forms a unique hydrogen bond with the chromophore, are required for far-red emission. Both the far-red emission and the time dependent shift in emission maximum are proposed to result from the interaction between the chromophore and Glu16. Our findings suggest that significant red shifts might be achieved in other fluorescent proteins using the strategy that led to the discovery of mPlum. [source] Crystal structure of the parasite inhibitor chagasin in complex with papain allows identification of structural requirements for broad reactivity and specificity determinants for target proteasesFEBS JOURNAL, Issue 3 2009Izabela Redzynia A complex of chagasin, a protein inhibitor from Trypanosoma cruzi, and papain, a classic family C1 cysteine protease, has been crystallized. Kinetic studies revealed that inactivation of papain by chagasin is very fast (kon = 1.5 × 106 m,1·s,1), and results in the formation of a very tight, reversible complex (Ki = 36 pm), with similar or better rate and equilibrium constants than those for cathepsins L and B. The high-resolution crystal structure shows an inhibitory wedge comprising three loops, which forms a number of contacts responsible for the high-affinity binding. Comparison with the structure of papain in complex with human cystatin B reveals that, despite entirely different folding, the two inhibitors utilize very similar atomic interactions, leading to essentially identical affinities for the enzyme. Comparisons of the chagasin,papain complex with high-resolution structures of chagasin in complexes with cathepsin L, cathepsin B and falcipain allowed the creation of a consensus map of the structural features that are important for efficient inhibition of papain-like enzymes. The comparisons also revealed a number of unique interactions that can be used to design enzyme-specific inhibitors. As papain exhibits high structural similarity to the catalytic domain of the T. cruzi enzyme cruzipain, the present chagasin,papain complex provides a reliable model of chagasin,cruzipain interactions. Such information, coupled with our identification of specificity-conferring interactions, should be important for the development of drugs for treatment of the devastating Chagas disease caused by this parasite. [source] TEXTURAL CHANGES IN CHOCOLATE CHARACTERIZED BY INSTRUMENTAL AND SENSORY TECHNIQUESJOURNAL OF TEXTURE STUDIES, Issue 4 2009LIA M. ANDRAE-NIGHTINGALE ABSTRACT Cocoa butter has a distinct texture due to unique interactions of polymorphic lipid structures. Part of chocolate's appeal is smooth mouthfeel; as fat or sugar bloom forms, textural change is perceived. Correlation of instrumental and sensory texture analysis has not been conducted in stored chocolate. The objective of this study was to analyze texture and color of dark and milk chocolate stored under conditions leading to fat and/or sugar bloom by instrumental and sensory measurements. Milk and dark chocolate was stored 5 weeks at various temperatures and relative humidity (RH), followed by instrumental and sensory texture analysis. All attributes, except springiness, were significantly affected by treatments. According to partial least squares linear regression, instrumental hardness, cohesiveness, chewiness and gumminess modeled sensory hardness. The 30.0C incubator experienced temperature fluctuations, resulting in severe fat bloom. Temperature fluctuations during storage had more influence on texture perception than storage at high temperatures or high RH. PRACTICAL APPLICATIONS This research serves as an initial study on textural aspects of chocolate quality upon storage that is the first report to correlate instrumental textural analysis of chocolate to sensory evaluation. Storage temperature and humidity of chocolate greatly impacts consumer texture perception, which is valuable information to small chocolate handlers and manufacturers who have noted to us that many of the larger companies may have this information , but it is not widely available. It also sets the stage for more detailed studies on texture and flavor of chocolate during storage. Although many storage studies on chocolate exist, those that intertwine studies of quality from both an instrumental and a sensory standpoint are lacking. [source] Winds of time and place: How context has affected a 50,year marriagePERSONAL RELATIONSHIPS, Issue 3 2003George Levinger To examine the effects of contexts on a relationship, we consider the case of our own 50,year marriage and its preliminaries. We employ a three,level conception of a couple's environment. The macrocontext refers to the prevailing cultural winds in a society that affect all its residents during any given historical era. The mesocontext pertains to the settings in which a particular relationship operates, such as its family and other social networks, physical habitats, work settings, or institutional associations, often chosen by the partners themselves. The microcontext is the pair's own intimate environment, constructed over time by the partners' unique interactions. Each of these contexts has affected us. We describe and analyze instances of luck, choice, and dyadic interaction in our 52,year relationship. [source] Covalent Attachment of Bacteriorhodopsin Monolayer to Bromo-terminated Solid Supports: Preparation, Characterization, and Protein StabilityCHEMISTRY - AN ASIAN JOURNAL, Issue 7 2008Yongdong Jin Dr. Abstract The interfacing of functional proteins with solid supports and the study of related protein-adsorption behavior are promising and important for potential device applications. In this study, we describe the preparation of bacteriorhodopsin (bR) monolayers on Br-terminated solid supports through covalent attachment. The bonding, by chemical reaction of the exposed free amine groups of bR with the pendant Br group of the chemically modified solid surface, was confirmed both by negative AFM results obtained when acetylated bR (instead of native bR) was used as a control and by weak bands observed at around 1610,cm,1 in the FTIR spectrum. The coverage of the resultant bR monolayer was significantly increased by changing the pH of the purple-membrane suspension from 9.2 to 6.8. Although bR, which is an exceptionally stable protein, showed a pronounced loss of its photoactivity in these bR monolayers, it retained full photoactivity after covalent binding to Br-terminated alkyls in solution. Several characterization methods, including atomic force microscopy (AFM), contact potential difference (CPD) measurements, and UV/Vis and Fourier transform infrared (FTIR) spectroscopy, verified that these bR monolayers behaved significantly different from native bR. Current,voltage (I,V) measurements (and optical absorption spectroscopy) suggest that the retinal chromophore is probably still present in the protein, whereas the UV/Vis spectrum suggests that it lacks the characteristic covalent protonated Schiff base linkage. This finding sheds light on the unique interactions of biomolecules with solid surfaces and may be significant for the design of protein-containing device structures. [source] | ||||||||||