RESUMEN
The anti-adhesive strategy, consisting of disrupting bacterial attachment to the host cells, is widely explored as an alternative to antibiotic therapies. Recently, thiazolylmannosides (TazMans) have been identified as strong anti-adhesives of E. coli strains implied in the gut inflammation of patients with Crohn's disease. In this work, we developed a second generation of TazMans with improved chemical stability. The anomeric nitrogen was substituted by short linkers and the compounds were assessed against the bacterial adhesin FimH and the clinically isolated LF82 E. coli strain in four in vitro assays. The results obtained on the FimH adhesin alone and the whole bacteria enabled the identification of a candidate for further in vivo evaluations.
Asunto(s)
Enfermedad de Crohn/tratamiento farmacológico , Escherichia coli/efectos de los fármacos , Proteínas Fimbrias/antagonistas & inhibidores , Manósidos/farmacología , Tiazoles/farmacología , Adhesinas de Escherichia coli , Enfermedad de Crohn/microbiología , Ensayo de Inmunoadsorción Enzimática , Escherichia coli/patogenicidad , Humanos , Espectroscopía de Resonancia Magnética , Manósidos/química , Pruebas de Sensibilidad Microbiana , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Tiazoles/químicaRESUMEN
Mono- and polyvalent ligands with strong affinities for the mannose-binding adhesin FimH were synthesised, and their anti-adhesive properties against ten E. coli strains were compared in two cell-based assays. The compounds were assessed against the non-pathogenic E. coli K12 and nine strains isolated by coproculture or from patients with osteoarticular infections (OIs), Crohn's disease (CD) and urinary tract infections (UTIs). The results showed that the compounds could inhibit the whole set of bacterial strains but with marked differences in terms of effective concentrations. The relative inhibitory potency of the monovalent compounds was also conserved for the ten strains and in the two assays. These results clearly suggest that a potent monovalent anti-adhesive assessed on a single E. coli strain will probably be effective on a broad range of strains and may treat diverse E. coli infections (OIs, CD and UTIs). In contrast, the polyvalent compounds showed a significant strain-dependancy in preventing E. coli attachment to intestinal cells. The multivalent antiadhesive effect may therefore vary depending on the E. coli strain tested.
Asunto(s)
Antibacterianos/química , Antibacterianos/farmacología , Adhesión Bacteriana/efectos de los fármacos , Infecciones por Escherichia coli/tratamiento farmacológico , Escherichia coli/efectos de los fármacos , Proteínas Fimbrias/antagonistas & inhibidores , Adhesinas de Escherichia coli/metabolismo , Escherichia coli/clasificación , Escherichia coli/metabolismo , Infecciones por Escherichia coli/microbiología , Proteínas Fimbrias/metabolismo , Humanos , LigandosRESUMEN
The aim of the present study was to further explore the influence of ingestion and mouth rinse with a carbohydrate-electrolyte solution (CES) on the performance during a approximately 1 h high-intensity time trial on trained subjects. Subjects rinsed around the mouth or ingested a 6% isotonic CES or placebo (14 mL/kg body weight) before and throughout a time trial in which they had to accomplish a set amount of work (975+/-85 kJ) as quickly as possible. In the mouth rinse conditions, time to complete the test was shorter (P=0.02) with CES (61.7+/-5.1 min) than with placebo (64.1+/-6.5 min), whereas in the ingestion conditions, there was no difference between placebo (62.5+/-6.9 min) and CES (63.2+/-6.9 min). Although power output and lactate concentration during exercise were significantly higher when subjects rinsed their mouth with CES compared with placebo, the rating of perceived exertion values did not differ. Blood glucose concentration increased after ingestion of but not after mouth rinse with CES. The interesting finding of the present study is that rinsing the mouth with but not ingestion of CES resulted in improved performance.
Asunto(s)
Carbohidratos de la Dieta/farmacología , Prueba de Esfuerzo/efectos de los fármacos , Ejercicio Físico/fisiología , Antisépticos Bucales , Adulto , Rendimiento Atlético/fisiología , Método Doble Ciego , Femenino , Glucosa/administración & dosificación , Humanos , Monosacáridos/administración & dosificación , Boca/fisiología , Antisépticos Bucales/farmacología , Consumo de Oxígeno/fisiología , Sacarosa/administración & dosificación , Edulcorantes/administración & dosificación , Factores de TiempoRESUMEN
Receptor-mediated uptake of orally administered antigen can lead to an antigen-specific immune response, whereas oral administration of most other non-replicating soluble antigens results in the induction of oral tolerance. In the present study, it is shown that fimbriae purified from an F4(K88)(+) enterotoxigenic Escherichia coli strain can function as a mucosal carrier molecule for the model antigen human serum albumin (HSA). Glutaraldehyde-coupled F4/HSA conjugates were able to bind F4 receptor positive (F4R(+)) enterocytes, but not to F4R(-) enterocytes. Moreover, oral immunization of F4R(+) pigs with F4/HSA conjugates induced a HSA-specific immune response, whereas oral immunization with HSA/HSA conjugates did not. This mucosal carrier function of F4 fimbriae was improved following oral co-administration of the F4/HSA conjugates with the mucosal adjuvant cholera toxin (CT) to F4R(+) pigs, since both humoral and cellular HSA-specific responses were significantly increased. In comparison with F4R(+) pigs, the HSA-specific response was reduced following oral F4/HSA+CT immunization of F4R(-) pigs. This indicates that F4 fimbriae as mucosal carrier and CT as adjuvant synergistically improve the induction of a HSA-specific immune response following oral immunization of pigs. These results could open new perspectives in the development of vaccines against enteropathogens.
Asunto(s)
Escherichia coli/inmunología , Fimbrias Bacterianas/inmunología , Albúmina Sérica/inmunología , Administración Oral , Animales , Anticuerpos Antibacterianos/sangre , Toxina del Cólera/inmunología , Inmunidad Mucosa , Inmunización , Inmunoglobulina A Secretora/biosíntesis , Activación de Linfocitos , PorcinosRESUMEN
This study investigated the effect of post-treatment phone contact on weight-loss maintenance and activity behaviour in obese youngsters. In all, 20 patients who completed a weight reduction program were randomly assigned to a 5-month maintenance programme (experimental) or control condition. Following the maintenance programme, patients sent a weekly activity diary to the therapist, who in turn phoned them biweekly to discuss their activities. Body weight, stature and physical activity were measured before and after the maintenance programme. The control group showed a continuous increase in overweight after initial treatment, while the experimental group showed a steep increase during the summer holidays (no intervention), but this increase slowed down during the maintenance programme (P<0.05). Moderate-to-high intensity activities increased during the maintenance programme in the experimental group, but decreased in the control group (P<0.001). In conclusion, post-treatment phone contact appears to have the potential to be an effective maintenance strategy in obese youngsters.
Asunto(s)
Obesidad/terapia , Cooperación del Paciente , Teléfono , Adolescente , Estatura/fisiología , Peso Corporal/fisiología , Ejercicio Físico/fisiología , Femenino , Humanos , Actividades Recreativas , Masculino , Obesidad/fisiopatología , Proyectos Piloto , Recurrencia , Estaciones del Año , Aumento de Peso/fisiologíaRESUMEN
We have previously observed that following the onset of moderate intensity cycle ergometry, the pulmonary O2 uptake (VO2) in trained cyclists often does not increase towards its steady-state value with the typical mono-exponential characteristics; rather, there is a transient "overshoot". The purpose of this study was to systematically examine this phenomenon by comparing the VO2 responses to two moderate-intensity work rates and one high-intensity work rate in trained and untrained subjects. Following a ramp exercise test to the limit of tolerance for the determination of the gas exchange threshold (GET) and VO2(peak), seven trained cyclists [mean (SD); VO2(peak) 66.6 (2.5) ml x kg(-1) x min(-1)] and eight sedentary subjects [VO2(peak) 42.9 (5.1) ml x kg(-1) x min(-1)] completed six step transitions from baseline cycling to work rates requiring 60% and 80% GET and three step transitions from baseline cycling to a work rate requiring 50% of the difference between GET and VO2(peak) (50%delta). VO2 was measured breath-by-breath and modelled using standard techniques. The sedentary subjects did not overshoot the steady-state VO2 at any intensity. At 60% GET, six of the seven cyclists overshot the steady-state VO2 [by an integral volume of 164 (44) ml between approximately 45 and 125 s]. At 80% GET, four of the seven cyclists overshot the steady-state VO2 [by an integral volume of 185 (92) ml between approximately 55 and 140 s]. None of the cyclists showed an overshoot at 50%delta. These results indicate that trained cyclists evidence an overshoot in VO2 before steady-state is reached in the transition to moderate-intensity exercise. The mechanism(s) responsible for this effect remains to be elucidated, as does whether the overshoot confers any functional or performance benefit to the trained cyclist.
Asunto(s)
Ejercicio Físico/fisiología , Oxígeno/sangre , Aptitud Física/fisiología , Adulto , Ciclismo/fisiología , Humanos , Masculino , Consumo de Oxígeno/fisiologíaRESUMEN
Previous studies have indicated that the (.-)VO(2) slow component is related to the recruitment of type II muscle fibres. We therefore hypothesised that an exercise and dietary regimen designed to deplete type I muscle fibres of glycogen would result in a greater contribution of type II muscle fibres to the exercise power output and therefore a larger amplitude of the (.-)VO(2) slow component. Eight male subjects took part in this study. On day 1, the subjects reported to the laboratory at 8 a.m., and completed a 9 min constant-load cycling test at a work rate equivalent to 85 % (.-)VO(2) peak. On day 2 at 12 p.m., the subjects were fed a 4200 kJ meal (60 % protein, 40 % fat); at 6 p.m. they completed a 2 h cycling test at 60 % (.-)VO(2) peak. On day 3 at 8 a.m., the subjects performed an exercise test identical to that of day 1. Metabolic and respiratory measurements indicated that our experimental design was effective in reducing the muscle glycogen content. (.-)VO(2) was significantly higher (by approximately 140 ml x min (-1)) throughout exercise following glycogen depletion but no appreciable changes in (.-)VO(2) kinetics were found: neither the time constant of the primary response (from 35.4 +/- 2.5 to 33.2 +/- 4.4 s) nor the amplitude of the slow component (from 404 +/- 95 to 376 +/- 81 ml x min (-1)) was significantly altered. Therefore, we suggest that the increased (.-)VO(2) throughout exercise and the unaltered (.-)VO(2) slow component following glycogen depletion might be explained by a shift towards a greater reliance on fat metabolism in type I muscle fibres with no appreciable change in fibre type recruitment patterns.
Asunto(s)
Ejercicio Físico/fisiología , Glucógeno/metabolismo , Fibras Musculares de Contracción Lenta/fisiología , Consumo de Oxígeno/fisiología , Adulto , Prueba de Esfuerzo , Humanos , Masculino , Fibras Musculares de Contracción Lenta/metabolismoRESUMEN
OBJECTIVE: (1) To examine the effects of attentional distraction on running time in an incremental treadmill test in obese youngsters; (2) to investigate whether distraction works at the same extent at the beginning and at the end of residential treatment; and (3) to explore the underlying mechanisms of the possible distraction effects. METHODS: Thirty severely obese youngsters (10 boys, 20 girls, age range 9-17) who were following a 10 month residential treatment, performed a treadmill test until exhaustion in four different sessions using a within subjects design. The two sessions at the beginning of the treatment and the two sessions at the end the treatment were counterbalanced, one with attentional distraction (music) and one without distraction. RESULTS: Obese youngsters ran significantly longer during distraction. This distraction effect seemed to be larger at the beginning compared to at the end of obesity treatment. The absence of differences between the condition with music and the condition without music on perceived bodily symptoms is in line with the idea that it took longer to perceive sufficient bodily sensations to decide to stop the treadmill test in the distraction condition. This interpretation is further corroborated by the physiological data indicating a superior peak performance in the condition with distraction. CONCLUSIONS: Attentional distraction has a positive effect on perseverance in obese youngsters. Further research has to show the usefulness of attentional distraction as a technique to increase exercise adoption and adherence in obesity treatment.
Asunto(s)
Tolerancia al Ejercicio , Música , Obesidad/terapia , Carrera/fisiología , Carrera/psicología , Adolescente , Atención , Actitud , Niño , Prueba de Esfuerzo/métodos , Femenino , Humanos , Masculino , Encuestas y Cuestionarios , Análisis y Desempeño de Tareas , Factores de Tiempo , Percepción del TiempoRESUMEN
In previous studies decreases in the VO(2) slow component were observed after prior heavy exercise. The observed effects after prior low-intensity exercise were rather controversial. The purpose of the present study was to more thoroughly examine the effects of prior low-intensity exercise on the VO(2) slow component. Furthermore, it has been suggested that the VO(2) slow component may be a determinant of exercise tolerance. Therefore we tested the hypothesis whether an attenuated VO(2) slow component induced by prior exercise could affect the time to exhaustion. Ten subjects performed four exercise protocols consisting of heavy cycling exercise (95 % VO(2)peak) until exhaustion. This constant-load exercise was performed without prior exercise (protocol NPE), or was preceded by 6 min heavy cycling exercise (protocol 6HPE), 12 min low-intensity cycling exercise (protocol 12LPE) or 6 min low-intensity cycling exercise (protocol 6LPE). The VO(2) slow component quantified as Delta VO(2 (end-2)) (669 +/- 90 ml x min (-1) in NPE) was significantly reduced after heavy as well as low-intensity exercise (respectively 47 %, 29 % and 17 % in 6HPE, 12LPE and 6LPE). This reduction lead to a significantly lower end VO(2) in 6HPE and 12LPE. The time to exhaustion (594 +/- 139 s in NPE), however, was unaffected by prior exercise rejecting our hypothesis that the attenuated VO(2) slow component could improve the capability to sustain exercise performance.
Asunto(s)
Adaptación Fisiológica/fisiología , Ciclismo/fisiología , Tolerancia al Ejercicio/fisiología , Ejercicio Físico/fisiología , Consumo de Oxígeno/fisiología , Adulto , Frecuencia Cardíaca/fisiología , Humanos , Ácido Láctico/sangre , Masculino , Valores de Referencia , TiempoRESUMEN
In previous studies conclusions about the effect of prior exercise on VO2 kinetics of subsequent high-intensity exercise are generally based on observed changes in the overall VO2 response without considering the effects on the VO2 fast and slow component. The aim of the present study was to examine the effect on the VO2 fast and slow component separately. Therefore 10 subjects performed an exercise protocol consisting of an initial 3 min period of unloaded cycling followed by two constant-load work bouts at a work rate corresponding to 90% VO2peak, separated by 3 min of rest and 3 min of unloaded cycling. VO2 was measured on a breath-by-breath basis, and the response curves were analysed by a biexponential model. To increase signal-to-noise ratio, subjects performed four repetitions of the exercise protocol, each separated by at least one day. There was no significant alteration in VO2 kinetic parameters of the primary, fast component after high-intensity exercise. However, there was a significant effect of prior high-intensity exercise on the VO2 kinetic parameters of the slow component. The time constant and the amplitude of the slow component were reduced by respectively 44% (from 231.0 +/- 111.7 s to 130.1 +/- 50.4 s) and 49% (from 824 +/- 270 ml x min(-1) to 417 +/- 134 ml x min(-1)). The results of this study indicate that the effect of high-intensity exercise on the VO2 kinetics of a subsequent high-intensity exercise is probably limited to an effect on the slow component.
Asunto(s)
Ejercicio Físico/fisiología , Consumo de Oxígeno , Resistencia Física , Adulto , Ciclismo/fisiología , Femenino , Humanos , Cinética , MasculinoRESUMEN
The crystal structures of cadmium/cadmium and zinc/calcium concanavalin A (con A) at pH 5.0 and pH 6.15, respectively, were determined. The structure of cadmium/cadmium con A confirms that the secondary Cd(2+)-binding site S3 is empty at pH 5. The metal-binding sites S1 and S2 are only very slightly affected by the substitution with cadmium. On the other hand, S1 and S2 and most of the protein surface of zinc/calcium con A at pH 6.15 differ from other fully metal-bound and carbohydrate-free structures. Most of these structural differences at the protein surface are a result of the interplay between metal binding, protonation and crystal packing. This interplay is expressed by relative rotations and translations of the con A units in alternative crystal packings and participation in space-group conversions inside crystals in situ. The particular crystal packing of zinc/calcium con A creates a novel zinc-binding site S4. The Zn(2+) ion in S4 ligates two aspartates from one tetramer and a histidine from a symmetry-related tetramer.
Asunto(s)
Cadmio/química , Calcio/química , Concanavalina A/química , Zinc/química , Ácido Aspártico/química , Sitios de Unión , Fabaceae/química , Histamina/química , Concentración de Iones de Hidrógeno , Modelos Moleculares , Lectinas de Plantas , Plantas Medicinales , Conformación Proteica , Difracción de Rayos XRESUMEN
The reversible binding of manganese and calcium to concanavalin A determines the carbohydrate binding of the lectin by inducing large conformational changes. These changes are governed by the isomerization of a non-proline peptide bond, Ala-207-Asp-208, positioned in a beta-strand in between the calcium binding site S2 and the carbohydrate specificity-determining loop. The replacement of calcium by manganese allowed us to investigate the structures of the carbohydrate binding, locked state and the inactive, unlocked state of concanavalin A, both with and without metal ions bound. Crystals of unlocked metal-free concanavalin A convert to the locked form with the binding of two Mn(2+) ions. Removal of these ions from the crystals traps metal-free concanavalin A in its locked state, a minority species in solution. The ligation of a metal ion in S2 to unlocked concanavalin A causes bending of the beta-strand foregoing the S2 ligand residues Asp-10 and Tyr-12. This bending disrupts conventional beta-sheet hydrogen bonding and forces the Thr-11 side chain against the Ala-207-Asp-208 peptide bond. The steric strain exerted by Thr-11 is presumed to drive the trans-to-cis isomerization. Upon isomerization, Asp-208 flips into its carbohydrate binding position, and the conformation of the carbohydrate specificity determining loop changes dramatically.
Asunto(s)
Concanavalina A/química , Concanavalina A/metabolismo , Calcio/metabolismo , Cristalografía por Rayos X , Electrones , Análisis de Fourier , Concentración de Iones de Hidrógeno , Isomerismo , Cinética , Ligandos , Manganeso/metabolismo , Modelos Químicos , Modelos Moleculares , Péptidos/metabolismo , Prolina/química , Unión Proteica , Conformación Proteica , Estructura Terciaria de ProteínaRESUMEN
The aim of the study was to examine to what extent prior high- or low-intensity cycling, yielding the same amount of external work, influenced the oxygen uptake (VO2) slow component of subsequent high-intensity cycling. The 12 subjects cycled in two protocols consisting of an initial 3 min period of unloaded cycling followed by two periods of constant-load exercise separated by 3 min of rest and 3 min of unloaded cycling. In protocol 1 both periods of exercise consisted of 6 min cycling at a work rate corresponding to 90% peak oxygen uptake (VO2peak). Protocol 2 differed from protocol 1 in that the first period of exercise consisted of a mean of 12.1 (SD 0.8) min cycling at a work rate corresponding to 50% VO2peak. The difference between the 3rd min VO2 and the end VO2 (deltaVO2(6-3)) was used as an index of the VO2 slow component. Prior high-intensity exercise significantly reduced deltaVO2(6-3). The deltaVO2(6-3) was also reduced by prior low-intensity exercise despite an unchanged plasma lactate concentration at the start of the second period of exercise. The reduction was more pronounced after prior high- than after prior low-intensity exercise (59% and 28%, respectively). The results of this study show that prior exercise of high as well as low intensity reduces the VO2 slow component and indicate that a metabolic acidosis is not a necessary condition to elicit a reduction in deltaVO2(6-3).
Asunto(s)
Ejercicio Físico/fisiología , Consumo de Oxígeno , Adulto , Ciclismo , Femenino , Humanos , Cinética , Ácido Láctico/sangre , Masculino , Factores de TiempoRESUMEN
Several novel structures of legume lectins have led to a thorough understanding of monosaccharide and oligosaccharide specificity, to the determination of novel and surprising quaternary structures and, most importantly, to the structural identification of the binding site for adenine and plant hormones. This deepening of our understanding of the structure/function relationships among the legume lectins is paralleled by advances in two other plant lectin families - the monocot lectins and the jacalin family. As the number of available crystal structures increases, more parallels between plant and animal lectins become apparent.
Asunto(s)
Carbohidratos/química , Lectinas/química , Sitios de Unión , Dimerización , Sustancias Macromoleculares , Modelos Moleculares , Reguladores del Crecimiento de las Plantas/química , Lectinas de Plantas , Plantas , Conformación Proteica , Estructura Cuaternaria de Proteína , Estructura Secundaria de ProteínaRESUMEN
The crystal structures of concanavalin A in complex with Man(alpha1-6)Man(alpha1-O)Me and Man(alpha1-3)Man(alpha1-O)Me were determined at resolutions of 2.0 and 2.8 A, respectively. In both structures, the O-1-linked mannose binds in the conserved monosaccharide-binding site. The O-3-linked mannose of Man(alpha1-3)Man(alpha1-O)Me binds in the hydrophobic subsite formed by Tyr-12, Tyr-100, and Leu-99. The shielding of a hydrophobic surface is consistent with the associated large heat capacity change. The O-6-linked mannose of Man(alpha1-6)Man(alpha1-O)Me binds in the same subsite formed by Tyr-12 and Asp-16 as the reducing mannose of the highly specific trimannose Man(alpha1-3)[Man(alpha1-6)]Man(alpha1-O)Me. However, it is much less tightly bound. Its O-2 hydroxyl makes no hydrogen bond with the conserved water 1. Water 1 is present in all the sugar-containing concanavalin A structures and increases the complementarity between the protein-binding surface and the sugar, but is not necessarily a hydrogen-bonding partner. A water analysis of the carbohydrate-binding site revealed a conserved water molecule replacing O-4 on the alpha1-3-linked arm of the trimannose. No such water is found for the reducing or O-6-linked mannose. Our data indicate that the central mannose of Man(alpha1-3)[Man(alpha1-6)]Man(alpha1-O)Me primarily functions as a hinge between the two outer subsites.
Asunto(s)
Concanavalina A/química , Disacáridos/química , Manósidos/química , Sitios de Unión , Conformación de Carbohidratos , Cristalografía por Rayos X , Modelos Moleculares , Unión Proteica , Termodinámica , Agua/químicaRESUMEN
We systematically analyzed the crystallographically determined water molecules of all known structures of RNase T1 and compared them to the ordered solvent in a large number of related microbial nucleases. To assess the crystallographers' impact on the interpretation of the solvent structure, we independently refined five validation structures from diffraction data derived from five isomorphous crystals of RNase T1. We also compared the positions of water molecules found in 11 published isomorphous RNase T1 inhibitor complexes. These data suggest that the positions of most of the waters located on the surface of a protein and that are well-determined in the experimental electron density maps are determined primarily by crystal packing forces. Water molecules with less well-defined electron density are in general unique to one or a small number of crystal structures. Only a small number of the well-defined waters are found to be independent of the crystal environment. These waters have a low accessible surface area and B-factor, and tend to be conserved in the crystal structures of a number of evolutionary related ribonucleases as well. A single water molecule is found conserved in all known microbial ribonucleases.
Asunto(s)
Ribonucleasas/química , Agua/química , Secuencia de Aminoácidos , Bacterias/enzimología , Cristalografía por Rayos X , Hongos/enzimología , Modelos Moleculares , Datos de Secuencia Molecular , Conformación Proteica , Proteínas Recombinantes/química , Homología de Secuencia de Aminoácido , SolventesRESUMEN
The seed lectin (DBL) from the leguminous plant Dolichos biflorus has a unique specificity among the members of the legume lectin family because of its high preference for GalNAc over Gal. In addition, precipitation of blood group A+H substance by DBL is slightly better inhibited by a blood group A trisaccharide (GalNAc(alpha1-3)[Fuc(alpha1-2)]Gal) containing pentasaccharide, and about 40 times better by the Forssman disaccharide (GalNAc(alpha1-3)GalNAc) than by GalNAc. We report the crystal structures of the DBL-blood group A trisaccharide complex and the DBL-Forssman disaccharide complex.A comparison with the binding sites of Gal-binding legume lectins indicates that the low affinity of DBL for Gal is due to the substitution of a conserved aromatic residue by an aliphatic residue (Leu127). Binding studies with a Leu127Phe mutant corroborate these conclusions. DBL has a higher affinity for GalNAc because the N-acetyl group compensates for the loss of aromatic stacking in DBL by making a hydrogen bond with the backbone amide group of Gly103 and a hydrophobic contact with the side-chains of Trp132 and Tyr104. Some legume lectins possess a hydrophobic binding site that binds adenine and adenine-derived plant hormones, i.e. cytokinins. The exact function of this binding site is unknown, but adenine/cytokinin-binding legume lectins might be involved in storage of plant hormones or plant growth regulation. The structures of DBL in complex with adenine and of the dimeric stem and leaf lectin (DB58) from the same plant provide the first structural data on these binding sites. Both oligomers possess an unusual architecture, featuring an alpha-helix sandwiched between two monomers. In both oligomers, this alpha-helix is directly involved in the formation of the hydrophobic binding site. DB58 adopts a novel quaternary structure, related to the quaternary structure of the DBL heterotetramer, and brings the number of know legume lectin dimer types to four.
Asunto(s)
Metabolismo de los Hidratos de Carbono , Lectinas/química , Oligosacáridos/química , Sistema del Grupo Sanguíneo ABO , Adenina/metabolismo , Sitios de Unión , Cristalografía por Rayos X , Antígeno de Forssman/metabolismo , Lectinas/genética , Modelos Moleculares , Mutagénesis Sitio-Dirigida , Oligosacáridos/metabolismo , Oligosacáridos de Cadena Ramificada , Lectinas de Plantas , Conformación Proteica , Rosales/química , Especificidad por SustratoRESUMEN
The aim of this study was to investigate the effects of exercise-induced and thermal dehydration on postural balance. Eight male subjects cycled for 2 h at a power output equal to 57-63% VO2max on two different occasions: once without drinking (NF) and once with intake of 1.9 l of a carbohydrate-electrolyte solution (FR). Before and after the exercise test, the velocity of the centre of pressure (COP) excursion was measured on a force platform during 30-s bipedal standing in normal position, feet side by side, and tandem position, feet heel to toe. On another occasion, eight subjects underwent seven consecutive sauna sessions (85 degrees C, 50% rh) of 15 min duration with no fluid replacement (S) to induce thermal dehydration. Mean fluid loss was 2.7 (+/- 0.4)%, 0.5 (+/- 0.5)% and 3.0 (+/- 0.6)% of body mass after NF, FR and S, respectively. Mean velocity of COP excursion after the exercise test was significantly higher in the NF than in the FR trial (p < 0.05). Postural stability was not influenced by S. In conclusion, prolonged exercise without fluid ingestion seems to negatively affect postural stability, whereas no effect is observed after exercise with fluid replacement or after thermal dehydration.
Asunto(s)
Deshidratación/fisiopatología , Ejercicio Físico/fisiología , Equilibrio Postural/fisiología , Postura/fisiología , Adulto , Análisis de Varianza , Deshidratación/complicaciones , Fluidoterapia , Calor/efectos adversos , Humanos , Masculino , Trastornos de la Sensación/etiologíaRESUMEN
The legume lectins are a large family of homologous carbohydrate binding proteins that are found mainly in the seeds of most legume plants. Despite their strong similarity on the level of their amino acid sequences and tertiary structures, their carbohydrate specificities and quaternary structures vary widely. In this review we will focus on the structural features of legume lectins and their complexes with carbohydrates. These will be discussed in the light of recent mutagenesis results when appropriate. Monosaccharide specificity seems to be achieved by the use of a conserved core of residues that hydrogen bond to the sugar, and a variable loop that determines the exact shape of the monosaccharide binding site. The higher affinity for particular oligosaccharides and monosaccharides containing a hydrophobic aglycon results mainly from a few distinct subsites next to the monosaccharide binding site. These subsites consist of a small number of variable residues and are found in both the mannose and galactose specificity groups. The quaternary structures of these proteins form the basis of a higher level of specificity, where the spacing between individual epitopes of multivalent carbohydrates becomes important. This results in homogeneous cross-linked lattices even in mixed precipitation systems, and is of relevance for their effects on the biological activities of cells such as mitogenic responses. Quaternary structure is also thought to play an important role in the high affinity interaction between some legume lectins and adenine and a series of adenine-derived plant hormones. The molecular basis of the variation in quaternary structure in this group of proteins is poorly understood.