Biomedical subjects

J Monreal

Publications and source records attributed to J Monreal.

At least 37 records · Page 2Linked to original sources

PubMed1988

Conformation of brain proteolipid apoprotein. Effects of sonication and n-octyl-beta-D-glucopyranoside detergent.

The conformation of brain proteolipid apoprotein (PLA) has been investigated using infrared spectroscopy and freeze-fracture electron microscopy. For this purpose, spectroscopic samples consisting of a mixture of liquid paraffin and wet protein have been prepared. These systems have allowed us to record the infrared spectra of PLA at neutral pH. The amide I and III regions reveal the existence of a predominantly alpha-helical structure, as well as the presence of minor beta-strands and random coil forms. The effect of sonication and a non-denaturing detergent, (n-octyl-beta-D-glucopyranoside), on the structure of the protein have also been investigated. Sonication produces an increase of the beta and unordered structures at the expense of the alpha-helical conformation. These structural changes are enhanced in the presence of the non-ionic detergent n-octyl-beta-D-glucopyranoside. Lipids protect the native protein structure from the effects of sonication. The aforementioned detergent changes the PLA conformation by increasing the alpha-helical content at the expense of beta-sheet and random coil forms. Therefore the PLA structure seems to be similar to the structures of other proteins intrinsic to non-neural membranes. The effects investigated also suggest that PLA behaves in a conformationally flexible manner.

Amides↗

PubMed1987

Vibrational spectra and structure of myelin membranes.

Raman and infrared spectroscopy have been simultaneously applied, for the first time, to the study of myelin membranes and their proteolipid protein (PLP) so as to obtain information on the secondary structure of proteins and the ordering of lipid chains. The vibrational spectra were recorded at physiological pH using a non-denaturing detergent (n-octyl-beta-D-glucopyranoside) in phosphate buffer. Neither the buffer nor the detergent interfere spectroscopically with the amide bands from proteins. The spectra reveal that the predominant secondary structure in the polypeptide backbone in myelin is the helix. The proteolipid protein was found to be more disordered than the polypeptide arrangement of the myelin membrane, as deduced from the relative intensities and halfwidths of characteristic infrared amide I bands. beta-form and turns are also present, the amount of these structures being higher in PLP. The study of the Raman spectra of vC-C and vC-H regions made it possible to obtain information on the lipid chain order.

Animals↗

PubMed1987

Ionophoric properties of the proteolipid apoprotein from bovine brain myelin.

Ionophoric properties of the Proteolipid Apoprotein have been assayed. This is a highly purified and delipidated intrinsic myelin membrane protein, isolated from bovine brain white matter. The preparation of myelin membrane vesicles or the incorporation of purified protein into Dimiristoylphosphatidylcholine liposomes have been carried out. According to our results, the myelin Proteolipid protein may act as a Na+ and Rb+ (K+) unidirectional ionophoretic channel, which main physiological role could be related to the maintenance of ionic equilibrium of myelin sheath around the axons.

Animals↗

PubMed1986

Freeze-fracture characterization of proteolipid protein and basic protein of central nervous system myelin incorporated in liposomes.

Proteolipid protein (PLP) and basic protein (BP) of central nervous system myelin were purified from calf brain white matter and incorporated in liposomes of L-dimyristoyl-alpha-phosphatidylcholine (DML) or in liposomes formed with an extract of natural lipids from myelin. Freeze-fracture replicas of the liposomes were prepared to study the number and size of intramembrane protein particles (IMP) in the fracture faces of the lipid bilayer. Globular and elongated IMP were observed in the freeze-fracture liposome membranes after incorporation of proteolipid protein. Globular IMP were the most frequently found (91-96% of the total IMP), and some of them showed a tiny black spot or pit on the top, suggesting the presence of hydrophilic channels in these particles. Globular and elongated IMP were also observed in the fractured membranes when basic protein was incorporated in liposomes. Again, globular IMP were the most frequent (92-95%) but no spots were present on the top. In addition, both globular and elongated IMP generated by basic protein were significantly larger than IMP generated by PLP. The proportion, size and form of globular and elongated particles generated by PLP and BP were unaffected by the amount of protein incorporated in liposomes (0.13-0.75 protein/lipid, w/w) nor by the type of lipid matrix used (DML or myelin natural lipid mixture). Intramembrane particles were absent from membranes of liposomes of pure lipid.

Animals↗

PubMed1986

Reversible thermal transition of brain myelin proteolipid. A preliminary report on a high-sensitivity differential scanning calorimetry study.

Brain myelin proteolipid has been investigated using high-sensitivity differential scanning calorimetry (DSC) under various conditions. Crude proteolipid with a 40% (w/w) content of protein gave rise to a reversible transition, centered at about 60 degrees C. The specific enthalpy of the transition was 50 +/- 5 J X g-1 with a calorimetric to van't Hoff enthalpy ratio of 5.7 +/- 0.5. To our knowledge this is the first intrinsic membrane protein in which a reversible thermal transition has been detected and investigated by DSC. Similar experiments were carried out using the recombinants of delipidated proteolipid and the pool of natural membrane lipids; in this case the transition was less enthalpic and showed lower cooperativity. The recombinants with lecithins, however, did not show any transition at 60 degrees C.

Animals↗

PubMed1984

The use of Moore swabs for isolation of Salmonella typhi from irrigation water in Santiago, Chile.

In Chile, a country with an exceedingly high incidence of typhoid, untreated sewage is applied directly to fields where salad vegetables are cultivated. Water used for irrigation was examined for the presence of Salmonella typhi, by making use of the sewer-swab technique. S typhi was isolated in 8 (11%) of 76 irrigation samples examined from nonindustrial, polluted water. This supports the hypothesis that crops grown with water contaminated with feces are important vehicles in the transmission of S typhi in this endemic area. Since sewage treatment plants will not be available in Santiago in the near future, emphasis is being placed on devising alternative methods of irrigation and on growing vegetables that are cooked before being eaten.

Bacteriological Techniques↗

PubMed1983

The delipidation of brain proteolipid protein by ultrafiltration.

It has been very difficult to prepare the apoprotein moiety of brain white matter proteolipid so that it is completely devoid of complex lipids, without suffering aggregation and protein denaturation. The reason is that complex lipids are tightly bound to the proteolipid apoprotein. Using a new ultrafiltration method, we obtained, in a gradual way and in a relatively short time, more than 99% delipidation in water-saturated n-butanol, with and without 0.1 M acetic acid, and recovered up to 86% of the protein with no detectable reducing sugars remaining. The delipidated protein remained in solution and in a relatively nondenatured state for several days. In 2% sodium dodecyl sulfate (SDS)-aqueous media, 90% of the lipids were removed and the yield of recovered protein in solution was near 90%; nearly 6% of the reducing sugars remained in the apoprotein. A higher delipidation was obtained by washing with 0.1 M NaOH. The content of reducing sugars was greater but the protein was less stable. When 10% SDS was employed to dissociate lipid-protein interaction, an almost complete delipidation was obtained and reducing sugars disappeared.

Animals↗

PubMed1982

Purification and chemical characterization of a W2 protein from brain myelin.

Starting from a pellet of beef brain myelin insoluble in chloroform/methanol (2:1, vol/vol)(Wolfgram protein fraction), a pure W2 protein with apparent molecular weight of 52,000 was isolated by a simple preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis method. A comparative chemical analysis was carried out between purified W2 and a standard tubulin. Glutamic acid and arginine were the N-terminals detected. Similar peptide maps and amino acid composition were also found in both proteins. Immunological cross-reactivity was detected when W2 protein was tested against antitubulin serum. These results suggest that W2 protein could have a tubulin-like protein nature that is associated with the myelin membrane and could play a role in the myelination process.

Amino Acids↗

PubMed1982

Method for lyophilizing brain proteolipid preparations that increases subsequent solubilization by detergents.

A frozen mixture of solubilized brain proteolipid proteins in chloroform-methanol is not sublimable in a vacuum. However, when 7 to 10 volumes of benzene were added to a chloroform-methanol solution containing 5 mg of proteolipid protein per ml, the proteolipid proteins remained in solution for a while and the frozen mixture was easily sublimated at 2mm Hg. Before the addition of benzene, higher concentrations of protein required the acidification of the medium to avoid precipitation of proteolipid proteins. In contrast to what happens when proteolipid proteins are obtained by the evaporation of the organic mixture at room temperature, the protein obtained by lyophilization was soluble in aqueous solutions of ionic and nonionic detergents. Sodium dodecyl sulfate at 0.6 to 0.7% concentration completely solubilized the proteolipid protein obtained by lyophilization. With the nonionic detergents Lubrol WX and Triton X-100, a solubilization between 50 and 65% was achieved. Sodium deoxycholate was practically ineffective. Triton X-100 showed selectivity in solubilizing certain proteins. The role of lipids in the solubilization of proteolipid proteins with detergents is discussed.

Animals↗

PubMed1976

The application of acetylated Sephadex for the separation of proteolipids.

The finding that acetylated Sephadex G 200 swells in chloroform/methanol mixture and still acts as a molecular sieve has permitted for the first time the separation of several proteolipid fractions from brain white matter, two of which appear almost free of phospholipids. This new procedure will be useful not only for a rapid purification of proteolipids and separation of lipids but also for general purposes of protein chemistry in organic media. However, it is predictable that one of the main uses of the method will be in the isolation of membrane carrier and of cholinergic receptor proteins, since these components appear to be of proteolipid(-like) nature.

Acetylation↗

PubMed1976