PubMed Health⌕ Search

Biomedical subjects

M Rojo

Publications and source records attributed to M Rojo.

At least 37 records · Page 2Linked to original sources
PubMed1997

Involvement of the transmembrane protein p23 in biosynthetic protein transport.

Here, we report the localization and characterization of BHKp23, a member of the p24 family of transmembrane proteins, in mammalian cells. We find that p23 is a major component of tubulovesicular membranes at the cis side of the Golgi complex (estimated density: 12,500 copies/micron2 membrane surface area, or approximately 30% of the total protein). Our data indicate that BHKp23-containing membranes are part of the cis-Golgi network/intermediate compartment. Using the G protein of vesicular stomatitis virus as a transmembrane cargo molecule, we find that p23 membranes are an obligatory station in forward biosynthetic membrane transport, but that p23 itself is absent from transport vesicles that carry the G protein to and beyond the Golgi complex. Our data show that p23 is not present to any significant extent in coat protein (COP) I-coated vesicles generated in vitro and does not colocalize with COP I buds and vesicles. Moreover, we find that p23 cytoplasmic domain is not involved in COP I membrane recruitment. Our data demonstrate that microinjected antibodies against the cytoplasmic tail of p23 inhibit G protein transport from the cis-Golgi network/ intermediate compartment to the cell surface, suggesting that p23 function is required for the transport of transmembrane cargo molecules. These observations together with the fact that p23 is a highly abundant component in the intermediate compartment, lead us to propose that p23 contributes to membrane structure, and that this contribution is necessary for efficient segregation and transport.

Amino Acid Sequence↗
PubMed1997

Generation of proteoliposomes from subcellular fractions.

Intracellular membranes are highly dynamic, yet they retain their identity and functional characteristics. Integral membrane proteins, which must confer this specific membrane identity, remain poorly characterized at the biochemical level, largely because detergent-mediated solubilization is required for purification and analysis, and several properties of integral membrane proteins can only be investigated when the molecule is properly embedded in a lipid bilayer. We present a method for the efficient reconstitution into proteoliposomes of integral membrane proteins from subcellular fractions. Integral membrane proteins were identified on high-resolution two-dimensional gels after selective extraction of soluble and peripheral membrane proteins; they accounted for 8% of the number of resolved polypeptides. A reconstitution procedure based on membrane solubilization with dodecyl-octaoxyethylene (C12E8) and subsequent detergent removal with BioBeads SM-2 resulted in the efficient reconstitution of several membrane proteins into proteoliposomes of uniform density. The generated proteoliposomes strongly resemble the starting membrane fraction in protein composition. This reconstitution allows the functional characterization of integral membrane proteins after enrichment and/or specific (immuno)depletion.

Animals↗
PubMed1997

Fal1p is an essential DEAD-box protein involved in 40S-ribosomal-subunit biogenesis in Saccharomyces cerevisiae.

A previously uncharacterized Saccharomyces cerevisiae gene, FAL1, was found by sequence comparison as a homolog of the eukaryotic translation initiation factor 4A (eIF4A). Fal1p has 55% identity and 73% similarity on the amino acid level to yeast eIF4A, the prototype of ATP-dependent RNA helicases of the DEAD-box protein family. Although clearly grouped in the eIF4A subfamily, the essential Fal1p displays a different subcellular function and localization. An HA epitope-tagged Fal1p is localized predominantly in the nucleolus. Polysome analyses in a temperature-sensitive fal1-1 mutant and a Fal1p-depleted strain reveal a decrease in the number of 40S ribosomal subunits. Furthermore, these strains are hypersensitive to the aminoglycoside antibiotics paromomycin and neomycin. Pulse-chase labeling of pre-rRNA and steady-state-level analysis of pre-rRNAs and mature rRNAs by Northern hybridization and primer extension in the Fal1p-depleted strain show that Fal1p is required for pre-rRNA processing at sites A0, A1, and A2. Consequently, depletion of Fal1p leads to decreased 18S rRNA levels and to an overall deficit in 40S ribosomal subunits. Together, these results implicate Fal1p in the 18S rRNA maturation pathway rather than in translation initiation.

Amino Acid Sequence↗
PubMed1996

[Prevalence of drinkers at risk and associated factors among men attending primary care clinics].

OBJECTIVES: To quantify the prevalence of risk drinkers among men seeking health care, establish consumption patterns and relate them to socio-demographic features and health habits. DESIGN: Crossover study. First phase of an experimental study. SETTING: Four primary care teams in Area 10, Madrid. PATIENTS: Males between 18 and 65 who attended for on-demand medical care. A systematic sample was selected (n = 562). MEASUREMENTS: A questionnaire collecting socio-demographic characteristics, health habits and alcohol consumption (frequency, consumption pattern, intake of Weekly Units of alcohol (WU) and alcohol-related problems). 94.2% replied. 38% were habitual drinkers (drink four or more days a week), while 18.9% were week-end drinkers. Prevalence of drinkers consuming over 21 WU was 24.8%; and over 35 WU, 16.2%. Whether the cut-off point was fixed at 21 or 35 WU. The alcohol taken was related to educational level, physical exercise, smoking and taking other drugs. On average, single people, smokers, other drug-consumers, ex-drinkers and habitual drinkers were those who displayed most problems connected with alcohol consumption. CONCLUSIONS: There is a similar profile of health habits for the groups consuming over 21 WU and 35 WU, which should make preventive care pay attention to both groups of drinkers.

Adolescent↗
PubMed1996

[Vitamin A, conjunctival cytology and clinical complications in children hospitalized with pneumonia].

To study the possible relationship between vitamin A status and the severity of pneumonia, an analysis was made of serum retinol levels, conjunctival cytology, and clinical severity of the disease in 178 children admitted to a pediatric hospital in Havana for acute infectious pneumonia. The pneumonia was considered to be complicated (n = 57) when the patient presented pleural discharge, pulmonary abscesses, or both; if not, it was considered uncomplicated (n = 121). Serum retinol values in both groups were similar and were not found to be associated with the severity of the pneumonia. On the other hand, children with alterations in conjunctival cytology were twice as likely to have complicated pneumonia as those with normal cytology (relative risk = 2.2, with a 95% confidence interval of 1.1 to 4.5).

Adolescent↗
PubMed1994

The mitochondrial ATP/ADP carrier: interaction with detergents and purification by a novel procedure.

The interaction of several classes of detergents with mitochondrial ATP/ADP carrier (AAC) was studied. The detergents that were best suited for solubilization of active AAC differed in several physico-chemical properties, but contained relatively rigid or planar hydrophobic (sub)moieties. Based on specific binding of AAC to Blue Sepharose, a novel method for the purification of the AAC was developed. The new method gave AAC samples which were devoid of non-essential lipids and allowed to purify AAC isoenzymes from several species and tissues to a significantly higher degree of purity than that achieved up to now. Western blot analysis of purified AACs with an antiserum against chicken heart AAC confirmed that immunological variability is more important between tissues than between species. In contrast to liver and kidney AACs, brain AAC displayed similar antigenic properties to heart AAC.

Animals↗
PubMed1994

Spatial arrangement of the heart muscle fascicles and intramyocardial connective tissue in the Spanish fighting bull (Bos taurus).

The spatial arrangement of the muscle fascicles and intramyocardial connective tissue was examined in the ventricles of the heart of the Spanish fighting bull (Bos taurus). In both ventricles, the muscle fascicles of the myocardium are arranged in 3 main directions, forming 3 muscle layers within the ventricular wall. The preferentially vertical arrangement of the muscle fascicles in the superficial and deep layers at the level of the fibrous aortic rings and the base of the semilunar valve leaflets suggests that these fascicles are actively involved in valvular dynamics. After controlled digestion of myocytes and elastic fibres with NaOH, a 3-dimensional arrangement of the scaffolding of connective tissue that supports the muscle fascicles and myocytes was observed. The arrangement and structure of this scaffolding may influence the order of contraction of muscle fascicles in different layers of the ventricle. In addition, differences were observed between the connective tissue scaffolding surrounding the myocytes of the 2 ventricles; these variations were correlated with the different biomechanical properties.

Animals↗
PubMed1991

Mitochondrial creatine kinase mediates contact formation between mitochondrial membranes.

Purified mitochondrial creatine kinase (Mi-CK) (EC 2.7.3.2) from chicken heart was shown to interact simultaneously with purified inner and outer mitochondrial membranes, thereby creating an intermembrane chondrial membranes, thereby creating an intermembrane were purified from rat liver and thus were fully devoid of Mi-CK. Intermembrane contact formation was demonstrated by measuring the binding of inner membrane vesicles to outer membranes spread at the air-water interface. Mi-CK also mediated intermembrane adhesion when membranes formed with total lipid extracts of both membranes were used, pointing to the role of lipids as potential membrane anchors of Mi-CK in the mitochondrial intermembrane space. Other enzymes of the intermembrane space that (like Mi-CK) are also cationic, as well as cytosolic isoenzymes of creatine kinase, failed to induce contact formation. Thus, of the proteins tested, membrane contact formation was specific for Mi-CK. The two oligomeric forms of Mi-CK (octamer and dimer) differed in their ability to mediate intermembrane adhesion, the octamer being more potent. Highly basic peptides, i.e. poly-L-lysines, were shown to strongly interact with membranes formed with lipid extracts of mitochondrial membranes: they both induced intermembrane binding and fusion. Interestingly, the extent of contact formation mediated by poly-L-lysines was lower than that of octameric Mi-CK. The implications of these findings on the function and localization of Mi-CK and on the structure of the mitochondrial intermembrane compartment are discussed.

Animals↗
PubMed1991

Interaction of mitochondrial creatine kinase with model membranes. A monolayer study.

The interaction of mitochondrial creatine kinase (Mi-CK; EC 2.7.3.2) with phospholipid monolayers and spread mitochondrial membranes at the air/water interface has been investigated. It appeared that Mi-CK penetrated into these monolayers as evidenced by an increase in surface pressure upon incorporation of Mi-CK. The increase in surface pressure was dependent on (1) the amount and (2) the oligomeric form of Mi-CK in the subphase, as well as on (3) the initial surface pressure and (4) the phospholipid composition of the monolayer. In this experimental system Mi-CK was able to interact equally well with both inner and outer mitochondrial membranes.

Animals↗
PubMed1990

The role of contact sites between inner and outer mitochondrial membrane in energy transfer.

Three functions have been suggested to be localized in contact sites between the inner and the outer membrane of mitochondria from mammalian cells: (i) transfer of energy from matrix to cytosol through the action of peripheral kinases; (ii) import of mitochondrial precursor proteins; and (iii) transfer of lipids between outer and inner membrane. In the contact site-related energy transfer a number of kinases localized in the periphery of the mitochondrion play a crucial role. Two examples of such kinases are relevant here: (i) hexokinase isoenzyme I which is capable of binding to the outer aspect of the outer membrane; and (ii) the mitochondrial isoenzyme of creatine kinase which is localized in the intermembrane space. Recently, evidence was presented that both hexokinase and creatine kinase are preferentially localized in contact sites (Adams, V. et al. (1989) Biochim. Biophys. Acta 981, 213-225). The aim of the present experiments was two-fold. First, to establish methods which enable the bioenergetic aspects of energy transfer mediated by kinases in contact sites to be measured. In these experiments emphasis was on hexokinase, while 31P-NMR was the major experimental technique. Second, we wanted to develop methods which can give insight into factors playing a role in the formation of contact sites involved in energy transfer. In the latter approach, mitochondrial creatine kinase was studied using monolayer techniques.

Animals↗
PubMed1989

Reproductive function and sexual behaviour in female rats exposed to immobilization stress or ACTH injections during gestation.

Pregnant primiparous rats were exposed to low immobilization stress or ACTH injections (one unit) throughout pregnancy. We measured the following parameters in the female offsprings: puberty (vaginal opening and first oestrous), oestrous cycle and sexual behaviour. Compared to the controls, female sexual receptivity measured by means of lordosis and the lordosis quotient (LQ) increased in both the experimental groups, but the puberty parameters of the offspring did not alter.

Adrenocorticotropic Hormone↗
PubMed1986

Prenatal stress: effects on sexual receptivity in female rats.

Pregnant primiparous rats were submitted to daily immobilization stress for 3 hour periods which were increased 15 minutes every third day. A considerable increase in the receptivity to the male was observed in the prenatally stressed females versus controls. Female offspring from both groups did not show any differences in body, adrenal, ovary and brain weights.

Animals↗
PubMed1985

Effects of low stress during pregnancy on certain parameters of the offspring.

In recent years the effect of intense stress applied during pregnancy has been studied. In this work, we exposed the mother to low immobilization stress throughout the entire pregnancy. This treatment that increased the sexual capacity in males had no detectable effects on the reproductive behavior of females. A positive role of this phenomenon in nature is suggested, as the offspring of stressed mothers would be more capable of facing difficult and selective environments.

Animals↗
PubMedDate not supplied

The structure of mitochondrial creatine kinase and its membrane binding properties.

The biochemical and biophysical characterization of the mitochondrial creatine kinase (Mi-CK) from chicken cardiac muscle is reviewed with emphasis on the structure of the octameric oligomer by electron microscopy and on its membrane binding properties. Information about shape, molecular symmetry and dimensions of the Mi-CK octamer, as obtained by different sample preparation techniques in combination with image processing methods, are compared. The organization of the four dimeric subunits into the Mi-CK complex as apparent as apparent in the end-on projections is discussed and the consistently observed high binding affinity of the four-fold symmetric end-on faces towards many support films and towards each other is outlined. A study on the oligomeric state of the enzyme in solution and in intact mitochondria, using chemical crosslinking reagents, is presented together with the results of a search for a possible linkage of Mi-CK with the adenine nucleotide translocator (ANT). The nature of Mi-CK binding to model membranes, demonstrating that rather the octameric than the dimeric subspecies is involved in lipid interaction and membrane contact formation, is resumed and put into relation to our structural observations. The findings are discussed in light of a possible in vivo function of the Mi-CK octamer bridging the gap between outer and inner mitochondrial membranes at the contact sites.

Animals↗
PubMedDate not supplied

Frequency of delta-F508 mutation and XV2C/KM19 haplotypes in Cuban cystic fibrosis families.

We tested the frequency of the delta F508 mutation and haplotypes linked to the cystic fibrosis (CF) gene in Cuba. The delta F508 deletion was detected in 34.0% of the CF chromosomes. There was a shortage of delta F508 heterozygotes, suggesting non-randomness in mating patterns. Haplotype B (XV2C/KM19 1/2) was found on 40.5% of the CF chromosomes (71.5% of delta F508 chromosomes, 28.3% of non-delta F508 CF chromosomes) against 13.5% of non-CF chromosomes.

Cuba↗