Cytoplasmic surface

The anion exchange protein (band 3) is a transmembrane glycoprotein, with its carboxyl terminal end on the external surface of the membrane and its amino terminal end on the cytoplasmic surface. It is an example of a multipass membrane protein, extending across the... 

More than half of the total mass of the ATPase molecule is exposed on the cytoplasmic surface of the membrane, forming the 40-A x 60-A particles seen by negative staining electron microscopy [88 93]. 

The assignment of the six additional transmembrane helices (M5-M10) is less conclusive. Since both the N-terminal and the C-terminal segments of the Ca " -ATPase molecule are exposed on the cytoplasmic surface of the membrane [112,135-138], the number of transmembrane sequences must be even, probably eight or ten odd numbered transmembrane domains (seven or nine) appear to be excluded [138,139]. 

The A20 antibody did not bind significantly to native SR vesicles, but solubilization of the membrane with C Eg or permeabilization of the vesicles by EGTA exposed its epitope and increased the binding more than 20-fold [139], By contrast, the A52 antibody reacted freely with the native sarcoplasmic reticulum, while the A25 antibody did not react either in the native or in the C Eg solubilized or permeabilized preparations, and required denaturation of Ca " -ATPase for reaction, Clarke et al, [139] concluded that the epitope for A52 is freely exposed on the cytoplasmic surface, while the epitope for A20 was assigned to the luminal surface, where it became accessible to cytoplasmic antibodies only after solubilization or permeabilization of the membrane. The epitope for A25 is assumed to be on the cytoplasmic surface in a folded structure and becomes accessible only after denaturation. 

Of the 20 residues that react with A-ethylmaleimide in the non-reduced denatured Ca -ATPase at least 15 are available for reaction with various SH reagents in the native enzyme [75,239,310]. These residues are all exposed on the cytoplasmic surface. After reaction of these SH groups with Hg-phenyl azoferritin, tightly packed ferritin particles can be seen by electron microscopy only on the outer surface of the sarcoplasmic reticulum vesicles [143,311-314]. Even after the vesicles were ruptured by sonication, aging, or exposure to distilled water, alkaline solutions or oleate, the asymmetric localization of the ferritin particles on the outer surface was preserved [311,313,314]. 

The Ca transport and Ca -stimulated ATPase activity of sarcoplasmic reticulum is inhibited by 10-30nmol dicyclohexylcarbodiimide per mg protein in a Ca free medium [372]. A23187 enhanced the sensitivity of the enzyme to DCCD, while Ca or Sr at micromolar concentrations prevented the inhibition. Since Ca -loaded vesicles retained their sensitivity to DCCD in a Ca -free medium, the reactivity of the enzyme with DCCD is controlled by the occupancy of the high-affinity Ca sites on the cytoplasmic surface of the membrane. 

Using these equations, Lowe and Walmsley [48] have calculated the dissociation constants for sugar binding at the extracellular surface of the membrane (K s = b a in Fig. 2) and at the cytoplasmic surface (K. = elf = bid) x [dgich]) from the estimated rate constants for carrier re-orientation and the measured Michaelis constants. The dissociation constant for binding at the extracellular surface of the membrane, calculated in this way, is approximately lOmM and is largely unaffec-... 

The most common second messenger activated by protein/peptide hormones and catecholamines is cyclic adenosine monophosphate (cAMP). The pathway by which cAMP is formed and alters cellular function is illustrated in Figure 10.1. The process begins when the hormone binds to its receptor. These receptors are quite large and span the plasma membrane. On the cytoplasmic surface of the membrane, the receptor is associated with a G protein that serves as the transducer molecule. In other words, the G protein acts as an intermediary between the receptor and the second messengers that will alter cellular activity. These proteins are referred to as G proteins because they bind with guanosine nucleotides. In an unstimulated cell, the inactive G protein binds guanosine diphosphate (GDP). When the hormone... 

Integral membrane proteins with one transmembrane domain may have soluble domains at either or both surfaces. An example of a monotopic protein, cytochrome b5 has a single hydrophobic segment that forms a hairpin loop, acting as an anchor to the cytoplasmic surface but probably not totally penetrating the bilayer. 

Schmidt-Lantermann clefts are structures where the cytoplasmic surfaces of the myelin sheath have not compacted to form the major dense line and therefore contain Schwann or glial cell cytoplasm (Fig. 4-9). They are common in peripheral myelin but rare in the CNS. These inclusions of cytoplasm are present in each layer of myelin. The clefts can be visualized in the unrolled myelin sheet as tubes of cytoplasm similar to the tubes making up the lateral loops but in the middle regions of the sheet, rather than at the edges (Fig. 4-9). 

Before myelination the axon lies in an invagination of the Schwann cell (Fig. 4-10A). The plasmalemma of the cell then surrounds the axon and joins to form a double membrane structure that communicates with the cell surface. This structure, called the mesaxon, then elongates around the axon in a spiral fashion (Fig. 4-10). Thus, formation of myelin topologically resembles rolling up a sleeping bag the mesaxon winds about the axon, and the cytoplasmic surfaces condense into a compact myelin sheath and form the major dense line. The two external surfaces form the myelin intraperiod line. 

The MBPs are extrinsic proteins localized exclusively at the cytoplasmic surface in the major dense line (Fig. 4-11), a conclusion based on their amino acid sequence, inaccessibility to surface probes and direct localization at the electron microscope level by immunocytochemistry. There is evidence to suggest that MBP forms dimers, and it is believed to be the principal protein stabilizing the major dense line of CNS myelin, possibly by interacting with negatively charged lipids. A severe hypomyelination and failure of compaction of the major dense line in MBP deficient shiverer mutants supports this hypothesis (Table 4-2). 

Myelin basic protein. In PNS myelin, MBP varies from approximately 5% to 18% of total protein, in contrast to the CNS, where it is close to 30% [ 1 ]. In rodents, the same four 21,18.5,17 and 14kDa MBPs found in the CNS are present in the PNS. In adult rodents, the 14kDa MBP is the most prominent component and is termed Pr in the PNS nomenclature. The 18.5 kDa component is present and is often referred to as the P, protein in the nomenclature of peripheral myelin proteins. Another species-specific variation in human PNS is that the major basic protein is not the 18.5 kDa isoform that is most prominent in the CNS but rather a form of about 17 kDa. It appears that MBP does not play as critical a role in myelin structure in the PNS as it does in the CNS. For example, the shiverer mutant mouse, which expresses no MBP (Table 4-2), has a greatly reduced amount of CNS myelin, with no compaction of the major dense line. By contrast, shiverer PNS has essentially normal myelin,both in amount and structure, despite the absence of MBP. This CNS/PNS difference in the role of MBP is probably because the cytoplasmic domain of P0 has an important role in stabilizing the major dense line of PNS myelin. Animals doubly deficient for P0 and MBP have a more severe defect in compaction of the PNS major dense line than P0-null mice, which indicates that both proteins contribute to compaction of the cytoplasmic surfaces in PNS myelin [23],... 

MBP) holds membranes together at the inner cytoplasmic surfaces in the CNS. Although MBP is also present in the PNS, it is much less abundant than in the CNS and its absence does not have a dramatic effect on PNS myelin compaction. 

Gumbiner, B. M. Proteins associated with the cytoplasmic surface of adhesion molecules. Neuron 11 551-564,1993. 

The interaction of IP3 with its receptor involves complex and poorly understood regulatory interactions among the receptor, IP3 and Ca2+ - the latter apparently exerting influence from both the cytoplasmic and luminal aspects of the receptor. Ca2+ in the lumen of the ER appears to sensitize the receptor to IP3. On the cytoplasmic surface, low concentrations of Ca2+ sensitize while higher concentrations are inhibitory. These actions may contribute to the all-or-none oscillatory behavior of [Ca2+] signals seen in some cell types. This phenomenon will be discussed in more detail below. 

Rhodopsin is a transmembrane protein linked to 11-c/s-retinal, which, on photoabsorption, decomposes to opsin and all-f/a/75-retinal. Rhodopsin has a molecular weight of about 40,000. Its C-terminus is exposed on the cytoplasmic surface of the disk, and its sugar-containing... 

Androutsellis-Theotokis, A., Ghassemi, F and Rudnick, G. (2001) A conformationally sensitive residue on the cytoplasmic surface of serotonin transporter. J. Biol. Chem. 9,9. 

Phospholipid turnover also takes place in an asymmetric manner. The enzymes responsible for phospholipid turnover in response to receptor-mediated phospholipase c activation are active from the cytoplasmic surface of the membrane. Likewise, diacylglycerol kinases converting the product of phospholipase c back into the key intermediate of phospholipid biosynthesis, phosphatidic acid, are also located on the cytoplasmic smface of the membrane (Sanjuan et al., 2001). 

Achve translocation of phospholipids aaoss the plasma membrane has been demonstrated both from the inner to the outer leaflet and from the outer to the inner leaflet. The translocation processes specifically transport phosphatidyserine and phosphatidylethanolamine from the cytoplasmic to the outer surface of the membrane while choline phosphatides are transported from the outer to the cytoplasmic surface. The rate of translocahon, in general, is greater for the amino phospholipids compared with the choline phospholipids. 

Kubo, S., Kitami, T., Noda, S., Shimura, H., Uchiyama, Y., et al. 2001. Parkin is associated with cytoplasmic surface of the synaptic vesicles. / Neurochem 77 1-13... 

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