Raw seeds of sixteen common bean (Phaseolus vulgaris L.) cultivars were evaluated along two consecutive harvests (2001/2002 and 2002/2003) concerning their physicochemical characteristics, as well as the effect of cooking and storage conditions on starch and dietary fiber contents. Serum lipids and blood glucose levels were also evaluated in normolipidemic-normoglycemic rats witch were fed diets containing bean cultivars with different soluble fiber/total fiber ratios (SF/TF): Pérola diet (0.11), Diamante Negro diet (0.19) and Iraí diet (0.26); control group was fed a standard diet (with insoluble fiber). Except for dry matter, moisture, and total dietary fiber, cultivars kept their chemical characteristics between harvests. Regarding similarity among macronutrient levels (crude protein-CP, total dietary fiber, insoluble dietary fiber, soluble dietary fiber-SF, digestible starch-DS, and resistant starch-RS) seeds from harvests 2001/2002 and 2002/2003 were categorized into four different groups; the same was done for micronutrients (Fe, Zn, Mn, Cu, Ca, Mg, and P), and four groups were also identified. Guateian 6662 and Rio Tibagi were considered the cultivars with the best nutritional profile (highest levels of CP, SF, DS, Fe, and Zn). Storage under refrigerated or freezing conditions did not change fiber content of cooked beans, but decreased their DS content and increased RS content, mainly in seeds with low RS levels before cooking. Concerning biological response, rats fed bean diets experienced lower values for serum cholesterol (P<0.05) and lower glycemic indexes (P<0.05). It was also observed a similar weight gain among groups, however animals fed bean based diets showed lower fat retention (P<0.05). The effects of bean diets on experimental groups were more remarkable in animals fed Iraí diet (SF/TF: 0.26). / Foram analisadas as características físico-químicas de sementes cruas de dezesseis cultivares de feijão comum (Phaseolus vulgaris L.), ao longo de duas safras consecutivas (2001/2002 e 2002/2003), bem como os teores de amido e fibra alimentar nas sementes após cozimento e estocagem. Avaliaram-se, também, os lipídeos séricos e a glicose sanguínea de ratos normolipidêmicos e normoglicêmicos, alimentados com dietas contendo cultivares de feijão com diferentes relações fibra solúvel/fibra total (FS/FT): dieta Pérola (0,11), dieta Diamante Negro (0,19) e dieta Iraí (0,26); o grupo controle recebeu dieta padrão (com fibra insolúvel). Exceto pelos teores de matéria seca, umidade e fibra alimentar total, as cultivares estudadas mantiveram suas características físico-químicas constantes ao longo das safras. Conforme a similaridade nos teores de macronutrientes (proteína bruta-PB, fibra alimentar total, fibra alimentar insolúvel, fibra alimentar solúvel-FS, amido disponível-AD e amido resistente-AR) as sementes das safras 2001/2002 e 2002/2003 foram categorizadas em quatro grupos distintos; da mesma forma, em relação aos micronutrientes (Fe, Zn, Mn, Cu, Ca, Mg e P), quatro grupos puderam ser identificados. As cultivares Guateian 6662 e Rio Tibagi apresentaram o melhor perfil nutricional (maiores teores de PB, FS, AD, Fe e Zn). A armazenagem sob refrigeração e o congelamento não determinaram alterações nos teores de fibra dos grãos cozidos, mas redução do AD e aumento do AR, principalmente naqueles com AR mais baixo antes do cozimento. Em relação à resposta biológica, os ratos alimentados com dietas contendo feijão apresentaram valores para colesterol sérico e índice glicêmico menores que os do grupo controle (p<0,05). Foi observado, também, que apesar do ganho de peso dos animais ter sido semelhante entre os grupos, os ratos alimentados com as dietas contendo feijão apresentaram menor retenção de gordura corporal (p<0,05). Os efeitos das dietas sobre os animais experimentais foram mais expressivos no grupo alimentado com a dieta Iraí (FS/FT: 0,26).

Les protéines Binder of Sperm (BSP) appartiennent à une superfamille de protéines exprimées dans le système reproducteur masculin, plus particulièrement dans les vésicules séminales chez les ongulés, et dans l’épididyme chez l’humain et la souris. Jusqu'à présent, des rôles variés chez différentes espèces ont été démontrés pour les protéines BSP, tels que dans la motilité et la capacitation chez le bovin. Cependant, leur rôle demeure élusif chez d’autres mammifères comme la souris et l’humain. Des études in vivo récentes ont démontré que la délétion des gènes Bsph1 et Bsph2 chez la souris n’a aucune conséquence sur la fertilité, et n’induit aucune anomalie au niveau de l'appareil reproducteur masculin. Afin d'élucider le rôle spécifique de la protéine BSP chez l'humain (BSPH1), nous avons d’abord développé une méthode de purification efficace permettant d’obtenir la protéine BSPH1 fonctionnelle car ces protéines ne sont présentes qu'en quantité infime dans l’épididyme humain. Suite, a la purification de BSPH1, j’ai réalisé des expériences in vitro et cherché à identifier son réseau d'interaction protéique. Il a été démontré que les protéines BSP interagissent avec des groupes pseudo-choline tels que le diéthylaminométhyle par affinité plutôt que par des interactions ioniques. Le diéthylaminoéthyle est chargé positivement et par conséquence, est un échangeur d'anions faible, mais les BSP interagissent avec affinité à la résine. Cette étude présente également une nouvelle méthode de purification rapide et peu coûteuse, qui fournit des protéines BSP recombinantes de grande pureté qui peuvent être utilisées pour étudier leurs rôles dans la fécondation chez les mammifères. Nous avons montré que la pré-incubation des ovocytes avec la protéine BSPH1 recombinante peut diminuer le taux de fécondation de manière dose-dépendante. Les spermatozoïdes ont également été pré-incubés avec un anticorps anti-BSPH1 et ont montré une diminution du taux de fécondation. Pour identifier le réseau d’interaction protéique de BSPH1, j'ai utilisé la méthode « Proximity-dependent biotin identification » (BioID) couplée à la spectrométrie de masse. Les résultats de la spectrométrie de masse ont démontré une interaction entre BSPH1 et toutes les sous-unités du complexe CCT / TRIC (Chaperonin containing tailless complex polypeptide 1 (CCT) ou tailless complex polypeptide 1 ring complex (TRiC)). Ce complexe interagit avec un autre complexe appelé BBSome (Bardet–Bied syndrome complex), qui joue un rôle important dans le transport de protéines à travers les cils primaires. BSPH1 a également interagi avec un grand nombre de protéines de la famille CEP (centrosome-associated proteins), importantes dans la formation des cils primaires par les microtubules et de la maturation du centrosome, qui soutiennent le rôle de BSPH1 dans les cils primaires. Dans l’ensemble, cette étude démontre que BSPH1 pourrait avoir un nouveau rôle en tant que chaperonne, à travers les cils primaires dans les cellules qui l’expriment dans l’appareil reproducteur masculin. / Binder of SPerm (BSP) proteins belong to a superfamily of proteins expressed in the male reproductive tract, particularly in seminal vesicles of ungulates (e.g., bovine, ram) and in the epididymis of humans and mice. So far, BSP proteins have been shown to play different roles in different species such as in motility and capacitation in bovine; however, their role remains unclear in other mammals. For instance, depletion of Bsph1/Bsph2 in mice had no effect on fertility. In order to elucidate the specific role of BSP protein in humans (BSPH1), I sought to investigate a purification method to produce functional human BSP protein, as these proteins are only present in minute amounts in the human epididymis. Following purification of BSPH1, I carried out in vitro experiments and sought to identify its protein interaction network. BSP proteins have been shown to interact with pseudo-choline groups such as diethylaminomethyl through affinity rather than ionic interactions. Diethylaminoethyl is positively charged and therefore is a weak anion exchanger, but BSPs interact through affinity to this resin. This study presents a new, rapid and cost-effective purification method that provides recombinant BSP proteins of a high purity level, which can be used to study their roles in mammalian fertilization. We showed that pre-incubation of oocytes with recombinant BSPH1 can decrease fertilization rate in a dose-dependant manner. Sperm were also preincubated with anti-BSPH1 antibody and showed a decrease in fertilization rate. Secondly, I used BioID (proximity-dependent biotin identification), coupled with mass spectrometry to identify the protein-protein interaction network of BSPH1 by proximity labeling. Mass spectrometry results showed an interaction between BSPH1 and all subunits of the CCT/TRIC complex (Chaperonin containing tailless complex polypeptide 1 (CCT) or tailless complex polypeptide 1 ring complex (TRiC). This complex interacts with another complex called BBSome (Bardet–Biedl syndrome complex), which plays a role in protein trafficking through primary cilium. I also identified BBS proteins, as well as other proteins, that interact with the BBSome complex and regulate protein trafficking in the cilia. BSPH1 also interacted with a large number of CEP (centrosome-associated proteins) family proteins, important in the formation of primary cilium through microtubules and centrosome maturation, which further support the potential implication of BSPH1 with the primary cilia. Overall, this study demonstrates that BSPH1 may have a new role as a chaperone involved in protein trafficking through the primary cilia in cells that express it in the male reproductive system