Cell structure Cytosol

The majority of CYP enzymes are located in a hydrophobic environment in the endoplasmic reticulum of cells, although cytosolic enzymes also exist, such as CYP101. In order to mimic the physiological environment of CYP enzymes, a number of groups have used phospholipids to construct biosensors such as DDAB, dimeristoyl-L-a-phosphatidylcholine (DMPC), dilauroylphosphatidylethanolamine (DLPE) and distearoylphosphatidylethanolamine (DSPE). Phospholipid layers form stable vesicular dispersions that bear structural relationship with the phospholipid components of biologically important membranes. By this way a membranous environment is created that facilitates electron transfer between the enzyme s redox center and the electrode. 

Recoverin is a Ca receptor with four EF structures and two Ca binding sites it can exist in the cytosol or associated with the membrane and has an N-terminal myristoyl residue as a lipid anchor. The distribution between free and membrane-associated forms is regulated by Ca. Binding of Ca to recoverin leads to its translocation from the cytosol to the membrane of the rod cells. Structural determination of recoverin in the Ca bound and Ca free forms (Ames et al., 1997) indicates that membrane association of recoverin is regulated by a Ca -myristoyl switch. The myristoyl residue can adopt two alternative positions in recoverin. In the absence of Ca, recoverin exists in a conformation in which the myristoyl residue is hidden in the iimer of the protein and is not available for membrane association. On Ca binding, a conformation change of recoverin takes place the myristoyl residue moves to the outside and can now associate with the membrane. 

More than 50 proteins have been discovered in the cytosol of nonmuscle cells that bind to actin and affect the assembly and disassembly of actin filaments or the cross-linking of actin filaments with each other, with other filamentous components of the cytoskeleton, or with the plasma membrane. Collectively, these are known as actin-binding proteins (ABPs). Their mechanisms of actions are complex and are subject to regulation by specific binding affinities to actin and other molecules, cooperation or competition with other ABPs, local changes in the concentrations of ions in the cytosol, and physical forces (Way and Weeds, 1990). Classifications of ABPs have been proposed that are based on their site of binding to actin and on their molecular structure and function (Pollard and Cooper, 1986 Herrmann, 1989 Pollard et al., 1994). These include the following ... 

The diversity of these subcellular actin structures is remarkable and appears to be determined by the interactions of many actin-binding proteins (ABPs) as well as by changes in the concentrations of intracellular signaling molecules such as Ca and cAMP, by small GTP-binding proteins, and by signals arising from mechanical stress. Approximately 50% of the actin molecules in most animal cells are unpolymerized subunits in the cytosolic pool and exist in a state of dynamic equilibrium with labile F-actin filamentous structures (i.e., new structures are formed while existing structures are renewed) (Hall, 1994). 

FIGURE 10.3 Structure of the BHTOH-KLH conjugate utilized for raising polyclonal antibodies. Western blots from 2D gels of cytosolic proteins isolated from mouse lung epithelial cells. The blot on the right is from cells incubated with BHT-QM and the blot on the left is from untreated cells. Immunoreactive proteins were identified by electrospray LC-MS analysis of the tryptic digests. Source From Ref. 39, with permission from the American Chemical Society. 

The basic characteristic of the membrane structure is its asymmetry, reflected not only in variously arranged proteins, but also in the fact that, for example, the outside of cytoplasmatic (cellular) membranes contains uncharged lecithin-type phospholipids, while the polar heads of strongly charged phospholipids are directed into the inside of the cell (into the cytosol). 

Not all the cellular DNA is in the nucleus some is found in the mitochondria. In addition, mitochondria contain RNA as well as several enzymes used for protein synthesis. Interestingly, mitochond-rial RNA and DNA bear a closer resemblance to the nucleic acid of bacterial cells than they do to animal cells. For example, the rather small DNA molecule of the mitochondrion is circular and does not form nucleosomes. Its information is contained in approximately 16,500 nucleotides that func-tion in the synthesis of two ribosomal and 22 transfer RNAs (tRNAs). In addition, mitochondrial DNA codes for the synthesis of 13 proteins, all components of the respiratory chain and the oxidative phosphorylation system. Still, mitochondrial DNA does not contain sufficient information for the synthesis of all mitochondrial proteins most are coded by nuclear genes. Most mitochondrial proteins are synthesized in the cytosol from nuclear-derived messenger RNAs (mRNAs) and then transported into the mito-chondria, where they contribute to both the structural and the functional elements of this organelle. Because mitochondria are inherited cytoplasmically, an individual does not necessarily receive mitochondrial nucleic acid equally from each parent. In fact, mito-chondria are inherited maternally. 

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