Transport mechanism, receptor/ligand binding

From studies of lipid-water mixtures and isolated membranes the general functional features of the bilayer are known barrier properties, lateral diffusion, acyl chain disorder and protein association. To vmderstand the mechanisms behind a wide spectrum of membrane functions, a detailed picture at the level of local curvature is needed. Examples are fusion processes, cooperativity in receptor/ligand binding or transport through the bilayer of the proteins that are constantly synthesised for export from the endoplasmic reticulum. Some preliminary discussions of the possibilities of curved, rather than flat, membremes follow. 

Recent evidence indicates that the 5-HT transporter is subject to post-translational regulatory changes in much the same way as neurotransmitter receptors (Blakeley et al. 1998). Protein kinase A and protein kinase C (PKC), at least, are known to be involved in this process. Phosphorylation of the transporter by PKC reduces the Fmax for 5-HT uptake and leads to sequestration of the transporter into the cell, suggesting that this enzyme has a key role in its intracellular trafficking. Since this phosphorylation is reduced when substrates that are themselves transported across the membrane bind to the transporter (e.g. 5-HT and fi -amphetamine), it seems that the transport of 5-HT is itself linked with the phosphorylation process. Possibly, this process serves as a homeostatic mechanism which ensures that the supply of functional transporters matches the demand for transmitter uptake. By contrast, ligands that are not transported (e.g. cocaine and the selective serotonin reuptake inhibitors (SSRIs)) prevent the inhibition of phosphorylation by transported ligands. Thus, such inhibitors would reduce 5-HT uptake both by their direct inhibition of the transporter and by disinhibition of its phosphorylation (Ramamoorthy and Blakely 1999). 

This lateral mobility within the membrane is important because it participates in the regulation of receptor binding to ligands. Receptor diffusivity determines the rate at which receptors can find each other, thereby determining the transport-limited rate of binding. Under purely diffusive mechanisms, the rate of receptor collision, k+ can be estimated according to the equation ... 

Pulmonary endotheUal cells are important in the regulation of circulating hormones (Ryan 1982), and consequently it is not surprising that certain xenobiotics, which have physicochemical properties similar to those of the endogenous substrates, also serve as substrates or ligands for the specialised enzymes, receptors, binding sites, and transport mechanisms localised on or in endothelial cells. 

The retrieval mechanism for the M6P receptor resembles the one previously described for the H/KDEL receptor [53]. Optimal binding of M6P receptor to M6P occurs at pH 6.5-6.7, the pH found in the TGN. When transport vesicles arrive at late endosomes, the pH is lowered by the action of H+ pumps. The affinity of the M6P receptor for its ligands is reduced at acid pHs, resulting in M6P receptor releasing the M6P in the late endosome. As a result, transport of lysosomal hydrolases occurs unidirectionally. Once the M6P receptor releases M6P-bearing hydrolases, the receptor can be returned to the TGN for reuse. Transport of the M6P receptor to either TGN or late endosome relies on signal peptides on the cytoplasmic tail region of the M6P receptor. 

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