Proteins visible

GFP Green fluorescent protein comes from the jellyfish Aequorea victoria. It fluoresces green, which is of puzzling utility to the jellyfish but of great use to scientists because it can act as a reporter molecule or can be used to make fusion proteins visible. Outdoing the jellyfish, molecular biologists have created GFP analogs that fluoresce in different colors (BFP, CFP, and YFP). 

Species origin tests, used to determine whether the specimen is human or from another source, are immunological in nature. Host animals, usually rabbits, are injected with protein from another species. The animal creates antibodies to the unknown material. Semm from the host animal, containing species (human, bovine, equine, canine, etc) specific antibodies, is tested against a dilute solution of blood (antigens) collected as evidence. A positive reaction is determined by a visible band where the antibodies and antigens come into contact. 

The transparency and refractive power of the lenses of our eyes depend on a smooth gradient of refractive index for visible light. This is achieved partly by a regular packing arrangement of the cells in the lens and partly by a smoothly changing concentration gradient of lens-specific proteins, the crystallins. 

Figure 8.3 The DNA-binding protein Cro from bacteriophage lambda contains 66 amino acid residues that fold into three a helices and three P strands, (a) A plot of the Ca positions of the first 62 residues of the polypeptide chain. The four C-terminal residues are not visible in the electron density map. (b) A schematic diagram of the subunit structure. a helices 2 and 3 that form the helix-turn-helix motif ate colored blue and red, respectively. The view is different from that in (a), [(a) Adapted from W.F. Anderson et al., Nature 290 754-758, 1981. (b) Adapted from D. Ohlendorf et al., /. Mol. Biol. 169 757-769, 1983.]...

This electron microscopy reconstruction has since been extended to high resolution (3 A) where the connections between the helices and the bound retinal molecule are visible together with the seven helices (Figure 12.3c). The helices are tilted by about 20° with respect to the plane of the membrane. This is the first example of a high-resolution three-dimensional protein structure determination using electron microscopy. The structure has subsequently been confirmed by x-ray crystallographic studies to 2 A resolution. 

Rossmann suggested that the canyons form the binding site for the rhi-novirus receptor on the surface of the host cells. The receptor for the major group of rhinoviruses is an adhesion protein known as lCAM-1. Cryoelectron microscopic studies have since shown that ICAM-1 indeed binds at the canyon site. Such electron micrographs of single virus particles have a low resolution and details are not visible. However, it is possible to model components, whose structure is known to high resolution, into the electron microscope pictures and in this way obtain rather detailed information, an approach pioneered in studies of muscle proteins as described in Chapter 14. 

During the separation itself some trouble can occur concerning the back pressure. An increasing back pressure indicates contamination of the column and thus should be monitored. If this happens or if a visible contamination of the sorbent is noticed, a regeneration of the column is necessary. However, if the back pressure rises very rapidly the column may be clogged by denatured proteins. As a first attempt, the frits should be replaced by new ones, trying the top adapter first. 

The cubic ternary complex model considers receptors and G-proteins as a synoptic system with some interactions that do not lead to visible activation. 

Constitutive receptor activity, receptors spontaneously produce conformations that activate G-proteins in the absence of agonists. This activity, referred to as constitutive activity, can be observed in systems in which the receptor expression levels are high and the resulting levels of spontaneously activating receptor species produce a visible physiological response. An inverse agonist reverses this constitutivie activity and thus reduces, in a dose-dependent... 

Just as myosins are able to move along microfilaments, there are motor proteins that move along microtubules. Microtubules, like microfilaments, are polar polymeric assemblies, but unlike actin-myosin interactions, microtubule-based motors exist that move along microtubules in either direction. A constant traffic of vesicles and organelles is visible in cultured cells especially using time-lapse photography. The larger part of this movement takes place on micrombules and is stimulated by phorbol ester (an activator of protein kinase C), and over-expression of N-J aj oncoprotein (Alexandrova et al., 1993). 

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