RESUMO
Alpha amylase inhibitor from Palo Fierro seeds (alphaAI-PF) was purified using affinity chromatography on a fetuin-fractogel column followed by anionic exchange chromatography. AlphaAI-PF has a molecular mass of 77kDa with two subunits (15.8 and 17.4 kDa), it is nonglycosylated and has pI of 4.7. AlphaAI-PF inhibited porcine pancreatic alpha-amylase (PPA) (1,4-alpha-D-glucan glucanohydrolase; EC 3.2.1.1), but was almost devoid of inhibitory activity on alpha-amylase extracts from Zabrotes subfasciatus (ZSA). Analysis of alphaAI-PF peptides showed a high homology to alphaAI-1 from Phaseolus vulgaris that also inhibits PPA.
Assuntos
Fabaceae/química , Sementes/química , alfa-Amilases/antagonistas & inibidores , Sequência de Aminoácidos , Relação Dose-Resposta a Droga , Dados de Sequência MolecularRESUMO
OBJECTIVES: The origin of the elevation of serum immunoglobulin A1 (IgA1) in Type 2 diabetes mellitus (DM) is unsettled. The aim of this study was to address the carbohydrate changes of serum IgA1 from patients with Type 2 diabetes mellitus, as a possible cause of the elevation. DESIGN AND METHODS: IgA1 was purified from sera of 6 DM patients and 4 healthy matched controls by using highly acetylated-Sepharose 6B, anti-IgA-agarose, and jacalin-agarose columns, and further separated into jacalin low-affinity, medium, and high-affinity fractions. Hinge and Fc fragments from native IgA1 were obtained and analyzed by using Sambucus nigra, Maackia amurensis, Arachis hypogaea, Erythrina cristagalli, and Ricinus communis lectins. RESULTS: The jacalin high-affinity fraction, mostly constituted by macromolecular IgA1, was more abundant in DM patients than in controls and also more reactive to Sambucus nigra, and Maackia amurensis lectins. CONCLUSIONS: Macromolecular serum IgA1 from DM patients is hypersialylated and this probably contributes to the high level of IgA1 in DM patients.
Assuntos
Diabetes Mellitus Tipo 2/sangue , Imunoglobulina A/sangue , Idoso , Carboidratos/sangue , Estudos de Casos e Controles , Cromatografia em Agarose , Eletroforese em Gel de Poliacrilamida , Feminino , Humanos , Imunoglobulina A/química , Imunoglobulina A/isolamento & purificação , Lectinas/metabolismo , Masculino , Pessoa de Meia-Idade , Ligação Proteica , Fatores de TempoRESUMO
One of the few sources of long-chain n-3 polyunsaturated fatty acids is fish oil, but considerable variation may exist according to species and season. In this study, the fatty acid profiles of sardine oils from Sardinops sagax caeruleus of the Gulf of California, Mexico, were evaluated in three seasonal catch periods. Oil quality was also evaluated by peroxide and free acid values. The most abundant fatty acids found in the oils were palmitic acid (19.3%), oleic acid (14.3%), eicosapentaenoic acid (EPA, 20.4%), and docosahexaenoic acid (DHA, 12.2%). There was no significant difference in the composition and quality among the six reduction plants where the samples were obtained. However, a significant difference in the proportion of EPA and DHA in one of the catch seasons analyzed was observed.
Assuntos
Ácidos Graxos/análise , Óleos de Peixe/química , Estações do Ano , Cromatografia Gasosa , Óleos de Peixe/normas , Controle de QualidadeRESUMO
1. Resolution of the fraction of sandbar shark (Carcharhinus plumbeus) serum that was soluble in 50% saturated ammonium sulfate by gel-immobilized metal-affinity chromatography allowed the isolation of a novel disulfide-bonded heterodimer of intact mass 70 kDa. 2. Following reduction, the molecule could be resolved into two chains of apparent mass 36 and 24 kDa. 3. The molecules were glycoproteins as determined by an observed reduction in molecular weight following enzymatic glycosylation. 4. The two separate chains were related to one another on the basis of amino-acid composition analysis and by comparison of the N-terminal amino acids (seven out of 10 identities). 5. The exact relationship of this molecule to characterized heterodimers of higher vertebrates is unknown. 6. Cross-linked agarose-acetate was synthesized and proved to be an efficient concentrating agent and also a hydrophobic interaction adsorbant.