52 research outputs found

    Interactions among the A and T Units of an ECF-Type Biotin Transporter Analyzed by Site-Specific Crosslinking

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    Energy-coupling factor (ECF) transporters are a huge group of micronutrient importers in prokaryotes. They are composed of a substrate-specific transmembrane protein (S component) and a module consisting of a moderately conserved transmembrane protein (T component) and two ABC ATPase domains (A components). Modules of A and T units may be dedicated to a specific S component or shared by many different S units in an organism. The mode of subunit interactions in ECF transporters is largely unknown. BioMNY, the focus of the present study, is a biotin transporter with a dedicated AT module. It consists of the S unit BioY, the A unit BioM and the T unit BioN. Like all T units, BioN contains two three-amino-acid signatures with a central Arg residue in a cytoplasmic helical region. Our previous work had demonstrated a central role of the two motifs in T units for stability and function of BioMNY and other ECF transporters. Here we show by site-specific crosslinking of pairs of mono-cysteine variants that the Ala-Arg-Ser and Ala-Arg-Gly signatures in BioN are coupling sites to the BioM ATPases. Analysis of 64 BioN-BioM pairs uncovered interactions of both signatures predominantly with a segment of ∼13 amino acid residues C-terminal of the Q loop of BioM. Our results further demonstrate that portions of all BioN variants with single Cys residues in the two signatures are crosslinked to homodimers. This finding may point to a dimeric architecture of the T unit in BioMNY complexes

    Relationship between volatile compounds and consumer-based sensory characteristics of bacon smoked with different brazilian woods

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    Bacon is a product made from pork meat that is subjected to curing, drying, and smoking. Researchers aim to associate the worldwide high-acceptance of such a product with the sensory and chemical properties of bacon. In this context, the objective of the present study was to characterize bacon samples smoked with different woods from reforestation using chemical and sensory methodologies, which were subsequently correlated by means of statistical multi-block analysis. Volatile compounds (VCs) of the smoked bacons were studied using solid-phase microextraction (SPME) coupled with gas chromatography-mass spectrometry (GC-MS), and the consumer sensory perception was explored by the Napping technique. VCs were identified in smoked samples, which triggered sensory attributes related to odor, flavor, and texture, such as "smoky flavor", "salty taste", "pleasant taste", "woodsy flavor", and "hard texture". Multi-block analysis showed that the sensory attributes were associated with a group of VCs and not by a single compound119839849FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2016/15012-2The authors are grateful to the São Paulo Research Foundation (FAPESP, Brazil) for funding the project No. 2016/15012-2. Erick Saldaña received the support of the “Consejo Nacional de Ciencia, Tecnología e Innovación Tecnológica - CONCYTEC” from Peru, in the framework of the CIENCIACTIVA programme, through the award of a PhD scholarship under contract agreement No. 104-2016-FONDECY
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