Cytosol entry

L-Ornithine transcarbamoylase catalyzes transfer of the carbamoyl group of carbamoyl phosphate to ornithine, forming citrulline and orthophosphate (reaction 2, Figure 29-9). While the reaction occurs in the mitochondrial matrix, both the formation of ornithine and the subsequent metabolism of citmlline take place in the cytosol. Entry of ornithine into mitochondria... 

Ramon G, Descombey P (1925) Sur 1 immunization antitetanique et sur la production de l antitoxine tetanique Compt Rend Soc Biol 93 508-98 Ratts R, Zeng H, Berg EA, Blue C, McComb ME et al. (2003) The cytosolic entry of diphtheria toxin catalytic domain requires a host cell cytosolic translocation factor complex. J Cell Biol 160 1139-50... 

Protein toxins acting intracellularly are often composed of two subunits (A/B model). One subunit is catalytic (A-subunit) and the other is responsible for binding and cell entry (B-subunit). Following binding to an extracellular membrane receptor, the toxins are endocytosed. From the endosomes, the A-subunit is directly (pH dqDendent) transferred into the cytosol (e.g., diphtheria toxin and anthrax toxin) or the toxin is transported in a retrograde manner via the golgi to the ER (e.g., cholera toxin), where translocation into the cytosol occurs [1]. 

Neutrophils represent an ideal system for studying osmotic effects on exocytosis. Stimulation of cytochalasin-B-treated neutrophils with the chemotactic peptide Jlf-formylmethionyl-leucyl-phenyl-alanine (FMLP) results in a rapid compound exocytosis up to 80% of lysosomal enzymes are released within 30 s (9-14). Secretion appears to be triggered by a rise in the level of cytosolic free calcium (15-18) promoted in part by entry of extracellular calcium through receptor-gated channels and in part by release of calcium that is sequestered or bound at some intracellular site (19-21). In this presentation, we augment our previously published data (22,23), which demonstrates that lysosomal enzyme release from neutrophils is inhibited under hyperosmotic conditions and that the rise in cytosolic calcium preceding secretion is inhibited as well. 

Figure 2. Reporting of cytosolic free calcium levels by indo-1. Increases in cytosolic calcium, due either to entry of extracellular calcium via calcium channels or to release of intracellular calcium sequestered in organelles such as smooth endoplasmic reticulum, results in formation of the indo-l-calcium complex. Fluorescence intensity at 400 nm (excitation at 340 nm) is proportional to the concentration of this complex the dissociation constant for this complex is about 250 nff (24), making this probe useful for detecting calcium activities in the range of 25 to 2500 nJ. ...

CICR). It is estimated that approximately 10% of the Ca involved in contraction enters the cytosol from the extracellular fluid and 90% from the sarcoplasmic reticulum. However, the former 10% is important, as the rate of increase of Ca in the myoplasm is important, and entry via the Ca channels contributes appreciably to this. 

Although the absence of paracellular transport across the BBB impedes the entry of small hydrophilic compounds into the brain, low-molecular-weight lipophilic substances may pass through the endothelial cell membranes and cytosol by passive diffusion [7]. While this physical barrier cannot protect the brain against chemicals, the metabolic barrier formed by the enzymes from the endothelial cell cytosol may transform these chemicals. Compounds transported through the BBB by carrier-mediated systems may also be metabolized. Thus, l-DOPA is transported through the BBB and then decarboxylated to dopamine by the aromatic amino acid decarboxylase [7]. 

Eiklid, K., Olsnes, S., and Pihl, A. (1980) Entry of lethal doses of abrin, ricin, and modeccin into the cytosol of Hela cells. Exp. Cell Res. 126, 321-326. 

Release is another area of difference conventional neurotransmitters are secreted from small synaptic vesicles (SSVs) after cytosolic [Ca2+] transiently reaches concentrations of 50-100 pmol/1, while peptides are released from LDCVs at lower concentrations of cytosolic [Ca2+] (Fig. 18-3). Conventional neurotransmitter release is thought to occur very close to the site of Ca2+ entry (see Chs 9,10), while neuropeptides are typically released at a distance from the site of Ca2+ entry (Fig. 18-4). Furthermore the Ca2+ that stimulates exocytosis from LDCVs may come from either internal stores or the extracellular fluid. Thus, the location of LDCVs relative to the site of Ca2+ influx has a very crucial impact on the intensity of stimulation necessary for secretion to occur. 

However, the current view of the regulation of calcium ion entry into the cytoplasm by PLC-linked stimuli holds that activation occurs not as a direct result of the action of IP3 on the plasma membrane but indirectly, as a result of depletion of calcium ions from an intracellular store by IP3 [14]. In the context of this capacitative model , the actions of intracellularly applied IP3 and heparin reflect the effects of these maneuvers on intracellular release process from ER into cytosol, rather than via the plasma membrane. The reported actions of I(1,3,4,5)P4, if in fact they do represent physiological control mechanisms, may reflect an ability of I(1,3,4,5)P4 to augment the calcium-releasing ability of IP3, rather than a distinct and... 

Diphtheria causes a demyelinative neuropathy. Coryne-bacterium diphtheriae colonizes the pharynx or open wounds, and secretes a protein exotoxin. The B subunit of this exotoxin binds to plasma membranes and facilitates entry into cytosol of the A subunit, which catalyzes ADP-ribosylation, and inactivation of an elongation factor required for protein synthesis. Cardiac muscle and Schwann cells are particularly susceptible to this toxin, and hence patients with diphtheria develop cardiomyopathy and demyelinative polyneuropathy [20]. While diphtheria is now uncommon because of childhood immunization against C. diphtheriae, the disruption in preventative medicine programs caused by disintegration of the Soviet Union was followed by a substantial incidence of diphtheritic polyneuropathy in Russia. 

In smooth muscle, just as in other cells, extracellular stimuli typically evoke an increase in cytosolic Ca2+ concentration ([Ca2+]c) by stimulating release of Ca2+ from intracellular stores or Ca2+ entry across the plasma membrane. Members of two closely related families of intracellular Ca2+ channels, ryanodine receptors and the receptors for inositol-1,4,5-trisphosphate (InsP3) appear to be expressed in all smooth muscle cells and they provide the most important route for release of Ca2+ from the sarcoplasmic reticulum (Sutko Airey 1996). The roles of additional Ca2+-mobilizing messengers, such as nicotinic acid adenine dinucleotide... 

Finally, we should mention the K+-dependent family of Na+/Ca2+ exchangers (NCKX), the first of which was discovered in retinal photoreceptors, which cotransport Ca2+ and K+ from the cytosol into the extracellular space, in exchange for the entry of Na+, with a stoichiometry of one K+ and one Ca2+ for four Na+. 

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