As lectinas são descritas como (glico)proteínas que se ligam, especificamente e reversivelmente, a carboidratos. Lectinas de leguminosas isoladas da subtribo Diocleinae (Canavalia, Dioclea eCratylia) são estruturalmente homólogas em relação às suas estruturas primárias. Demonstrou-se que as lectinas de DiocleinaeCanavalia brasiliensis, Dioclea guianensis eCanavalia ensiformis alteram diferentemente parâmetros fisiológicos em rins isolados de ratos. Dessa maneira, o objetivo deste estudo foi investigar o papel da lectina de Cratylia floribunda (CFL) na hemodinâmica renal e no transporte de íons em ratos. Em rins isolados perfundidos, CFL (10 mg/mL, n=5) aumentou a pressão de perfusão renal, a resistência vascular renal e reduziu o percentual do transporte tubular de K+, mas não alterou o fluxo urinário, a taxa de filtração glomerular e o percentual de transporte tubular dos íons sódio e cloreto. No leito mesentérico isolado perfundido, CFL (3 e 10 mg/mL/min, n=4) não alterou o tônus basal ou a contração do tecido induzida por fenilefrina (1 mM/mL/min). Em conclusão, a lectina de sementes de Cratylia floribunda altera parâmetros hemodinâmicos renais, provavelmente de origem tubular, e não por alterações hemodinâmicas.

Lectins have been described as glycoproteins that reversibly and specifically bind to carbohydrates. Legume lectins isolated from the subtribe Diocleinae (Canavalia, Dioclea andCratylia) are structurally homologous with respect to their primary structures. The Diocleinae lectins of Canavalia brasiliensis, Dioclea guianensis andCanavalia ensiformis have been shown to distinctly alter physiological parameters in isolated rat kidneys. Thus, the aim of this study was to investigate the effect of Cratylia floribunda lectin (CFL) on renal hemodynamics and ion transport in rats. In isolated perfused kidneys, CFL (10 mg/mL, n=5) increased RPP, RVR and decreased %TK+, but did not change urinary flow, glomerular filtration rate, sodium or chloride tubular transport. In isolated perfused mesenteric bed, CFL (3 and 10 mg/mL/min; n=4) did not alter tissue basal tonus or tissue contraction by phenylephrine (1 mM/mL/min). In conclusion, the seed lectin of Cratylia floribunda increased renal hemodynamic parameters showing a kaliuretic effect. This effect could be of tubular origin, rather than a result from haemodynamic alterations.

Lectins are carbohydrate-binding proteins of non-imune origin. This group of proteins is distributed widely in nature and they have been found in viruses, microorganisms, plants and animals. Lectins of plants have been isolated and characterized according to their chemical, physical-chemical, structural and biological properties. Among their biological activities, we can stress its fungicidal action. It has been previously described the effect of the lectins Dviol, DRL, ConBr and LSL obtained from the seeds of leguminous plants on the growth of yeasts isolated from vaginal secretions. In the present work the experiments were carried out in microtiter plates and the results interpreted by both methods: visual observations and a microplate reader at 530nm. The lectin concentrations varied from 0.5 to 256µg/mL, and the inoculum was established between 65-70% of trammitance. All yeast samples isolated from vaginal secretion were evaluated taxonomically, where were observed macroscopic and microscopic characteristics to each species. The LSL lectin did not demonstrate any antifungal activity to any isolate studied. The other lectins DRL, ConBr and DvioL, showed antifungal potential against yeast isolated from vaginal secretion. These findings offering offer a promising field of investigation to develop new therapeutic strategies against vaginal yeast infections, collaborating to improve women's health.

O objetivo deste estudo foi determinar o perfil de proteases em lesões cutâneas experimentais tratadas com a lectina isolada das sementes da Canavalia brasiliensis (ConBr) livre e conjugada com o seu açúcar específico. Lesões cirúrgicas foram produzidas assepticamente na região dorsal de camundongos (n=120), divididos de acordo com o tratamento empregado: Grupo NaCl (NaCl 150mM), Grupo manose (manose 100mM), Grupo ConBr (ConBr 100µg mL-1) e Grupo ConBr/manose (solução contendo ConBr 100µg mL-1 preparada em manose 100mM). Amostras da área lesada foram coletadas para determinação do perfil de proteases e atividade colagenolítica no 2°, no 7° e no 12° dia de pós-operatório. O perfil das proteínas realizado através de eletroforese SDS-PAGE demonstrou a presença de proteínas com massa molecular de 67kDa em todos os grupos. O Grupo ConBr/manose apresentou a maior atividade colagenolítica no 12° dia de pós-operatório. A lectina isolada das sementes da Canavalia brasiliensis influenciou a expressão de proteases com atividade colagenolítica podendo assim interferir no processo cicatricial das lesões cutâneas em camundongos.

The objective of the present study was determining the proteases profile of cutaneous healings treated with free and conjugated lectin of Canavalia brasiliensis (ConBr) and their specific sugar. An aseptic wound was produced in the thoracic area of the mice (n=120), divided according to the employed treatment: NaCl Group (150mM NaCl), manose Group (100mM manose), ConBr Group (100µg mL-1 ConBr) and ConBr/manose Group (solution containing 100µg mL-1 ConBr prepared in 100mM manose). Samples of the injured area were collected for determination of proteases profile and collagenolytic activity on 2nd, 7th e 12th days after the surgery. Electrophoresis SDS-PAGE demonstrated proteins with molecular mass of 67kDa in all groups. Group IV presented the highest collagenolytic activity on the 12th day post surgery. ConBr lectin influenced proteases expression with collagenolytic activity thus being able to intervene on skin wound healing in mice.

SUMMARY Biochemical and chemical partial characterization of Bauhinia forficata Link Seeds. Seeds of Bauhinia forficata species were submited to biochemical characterization concerning fatty acids analysis, protein fractionization, and hemaglutinanting activity. The seed elementary analysis showed a high protein and lipids contents with 21.24 % and 19.45 % respectively. The more abundant fatty acid was linoleic acid with 46.47 % of the lipidic fraction. With the exception of prolamins, the different proteic fractions (albumin, globulins, acid and basic glutelins) showed hemaglutinanting activity against rabbit red cells no treated and treated with proteolitic enzymes. The fraction acid glutelin showed the higher specific hemaglutinanting activity (1072.25 UH/mg P) against rabbit blood pre-treated with trypsin. Glutamin (16.20%) and Valin (11.07%) were the more abundant amino acids in the seeds. Therefore, B. forficata represent a possible optional source of food because exhibit a high energetic values.

Sementes quiescentes de Bauhinia forficata foram submetidas à caracterização bioquímica por meio de análise de ácidos graxos, fracionamento de proteínas e atividade hemaglutinante específica. A análise elementar da semente mostrou grande quantidade de proteína total e de lipídeos com 21,24 % e 19,45%, respectivamente. Na fração lipídica, o ácido linolêico foi o mais abundante com 46,47%. Com exceção das prolaminas, as diferentes frações protéicas (albuminas, globulinas, glutelinas ácidas e básicas) apresentaram atividade hemaglutinante contra hemácias de coelho tratadas e não tratadas com enzimas proteolíticas. A maior atividade hemaglutinante específica foi evidenciada na fração glutelinas ácidas (1.072,25 U.H./mg P) contra sangue de coelho tratado com tripsina. Os aminoácidos presentes em maior teor foram glutamina (16,20%) e valina (11,07%). Assim, por apresentarem alto valor energético as sementes de Bauhinia forficata são uma possível fonte opcional na alimentação.

A mobilização das proteínas de sementes de Pisum arvense L. durante a germinação na ausência de luz foi estudada. As sementes foram completamente exauridas de suas reservas após 22 dias de germinação e as plântulas cresceram até o 18° dia de germinação. Experimentos de eletroforese em gel de poliacrilamida em presença de SDS indicaram que as principais bandas protéicas, correspondendo às proteínas de reserva de alta massa molecular, foram detectadas apenas até o 7° dia após a germinação. Entretanto, as bandas protéicas correspondentes às lectinas da semente puderam ser detectadas ao longo de todo o processo germinativo, sugerindo que essas proteínas são fortemente resistentes à ação das proteases da semente e devem, de alguma forma, ser importantes para o estabelecimento das plântulas. A atividade hemaglutinante nos cotilédones foi detectada até o 22° dia de germinação, indicando que as lectinas também permaneceram ativas quando os cotilédones estavam bastante exauridos de suas reservas.

ABSTRACT - The mobilization of seed proteins from Pisum arvense L. during germination in the absence of light was studied. The seeds were found to be completely consumed 22 days after germination and seedlings ceased growth after the 18th day. SDS-PAGE indicated that the main protein bands correspond to high molecular mass storage proteins which undergo proteolysis in the initial stages of germination and are not detected after the 7th day of germination. However, the corresponding lectin profiles were detected during the entire germination process, suggesting that these proteins are strongly resistant to seed proteolytic enzymes and should be important for seedling establishment. Furthermore, haemagglutinating activity in cotyledons was detected until 22 days after germination, indicating that the lectins remain active even in senescent cotyledons.

As lectinas, proteinas que especificamente reconhecem estruturas que contém carboidratos, têm sido extensivamente estudadas em muitos aspectos bioquímicos e estruturais, objetivando estabelecer as bases moleculares deste evento não-catalítico. Por outro lado, os potenciais clínicos e agriculturais destas proteínas têm crescido rapidamente. Embora as lectinas, principalmente aquelas de legumes tenham sido bastante investigadas em suas propriedades biológicas, estudos sobre as funcões fisiológicas de lectinas são escassos na literatura. Além disto, a despeito da quantidade de dados acumulados sobre lectinas (como proteínas), o papel desempenhado por estas moléculas de sinalização é pobremente discutido. Valendo-se de nossos estudos sobre lectinas de leguminosas, principalmente da sub-tribo Diocleinae, e outros dados presentes na literatura, discutimos aqui, as principais hipóteses de suas funções com base na especificidade por carboidratos e glicanos complexos.

The lectins, proteins which specifically recognize carbohydrate moieties, have been extensively studied in many biochemical and structural aspects in order to establish the molecular basis of this non-catalytic event. On the other hand, their clinical and agricultural potentials have been growing fast. Although lectins, mainly those from legume plants, had been investigated for biological properties, studies about the physiological functions of lectins are scarce in literature. Therefore, despite the accumulated data on lectins (as proteins), the role played by these signalizing molecules is poorly discussed. In the light of our accumulated results on legume lectins, specially those obtained from plants belonging to the Diocleinae sub-tribe and available data in literature, we discuss here the main hypothesis of their functions according to their carbohydrate/glycan-binding specificity.

The carbohydrate-binding specificity of lectins from the seeds of Canavalia maritima and Dioclea grandiflora was studied by hapten-inhibition of haemagglutination using various sugars and sugar derivatives as inhibitors, including N-acetylneuraminic acid and N-acetylmuramic acid. Despite some discrepancies, both lectins exhibited a very similar carbohydrate-binding specificity as previously reported for other lectins from Diocleinae (tribe Phaseoleae, sub-tribe Diocleinae). Accordingly, both lectins exhibited almost identical hydropathic profiles and their three-dimensional models built up from the atomic coordinates of ConA looked very similar. However, docking experiments of glucose and mannose in their monosaccharide-binding sites, by comparison with the ConA-mannose complex used as a model, revealed conformational changes in side chains of the amino acid residues involved in the binding of monosaccharides. These results fully agree with crystallographic data showing that binding of specific ligands to ConA requires conformational chances of its monosaccharide-binding site.

Lectins, carbohydrate-binding proteins of non-immune origin, that agglutinate cells or precipitate polysaccharides and glycoconjugates, are well distributed in nature, mainly in the Plant Kingdom. The great majority of the plante lectins are present in seed cotyledons where they are found in the cytoplasm or int he protein bodies, although they have also been found in roots, stems and leaves. Due to their peculiar properties, the lectins are used as a tool both for analytical and preparative purposes in biochemistry, cellular biology, immunology and related areas. In agriculture and medicine the use of lectins greatly improved in the last few years. The lextins, with few exceptions, are glycoproteins, need divalent cations to display full activity and are, in general, oligomers with variable molecular weight. Although the studies on lectins have completed a century, their role in nature is yet ynknown . Several hypotheses on their physiological functions have been suggested. Thus, lectins could play important roles in defense against pathogens, plant-microorganism symbiosis, cell organization, embryo morphogenesis, phagocytosis, cell wall elongation, pollen recognition and as reserve proteins. A brief review on the general properties and roles of the lectins is given.

A comparação da metabolização da lectina de Canavalia brasiliensis com as demais proteínas de reservas durante a germinação das sementes na presença de luz foi acompanhada através da combinação de dosagem de proteínas solúveis, determinação da atividade hemaglutinante, cromatografia de afinidade em coluna de Sephadex G-50 e eletroforese em gel de poliacrilamida em presença de dodecilsulfato de sódio (SDS) e 2-mercaptoetanol. Os resultados foram comparados com resultados anteriormente obtidos com sementes germinadas na ausência da luz e ficou evidenciado que, mesmo com as plântuias fotossintetizando, a lectina de Canavalia brasiliensis é preservada, sendo metabolizada após as>demais proteínas de reserva. Estes dados sugerem também que estas proteínas devam ter funções específicas durante o ciclo vital da planta e não se constituem apenas em proteínas de reserva.

Canavalia brasiliensis seed lectin during germination in the presence of light. The mobilization of the lectin of Canavalia brasiliensis during seed germination, in the presence of light, was studied in comparison with the reserve proteins. Methods employed were determination of soluble protein content, hemagglutinating activity, affinity chromatography on Sephadex G-50 column, and PAGE-SDS. The results showed that the lectin was metabolized later than the reserve proteins suggesting that it may have a special function other than a simple reserve protein.

Eletroforese em gel de poliacrilamida das proteínas solúveis extraídas de sementes de doze espécies da tribo Diocleae, apresenta padrão qualitativo semelhante no que diz respeito às bandas correspondentes às lectinas, sugerindo a possibilidade de seu uso como marcadores taxonômicos.

Polyacrylamide gel electrophoresis of the soluble proteins extracted from seeds of twelve species belonging to the tribe Diocleae showed substantive qualitative pattern similiarities between the bands correspondent to the lectins.