The zymogen‐enteropeptidase system: A practical approach to study the regulation of enzyme activity by proteolytic cleavage*

Pizauro, João Martins; Ferro, J. A.; Lima, Andréa Cristina Frizzas de; Routman, Karina S.; Portella, Maria Célia

doi:10.1002/bmb.2004.494032010311

Cited by 5 publications

(2 citation statements)

References 9 publications

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“…Fast and efficient analysis of such compounds is required in molecular biology and biochemistry [3,4]. Here, biosensors are the most promising tools since they do not require sophisticated sample preparation, are simple to use, and can be successfully applied in biological and biochemical education [5].…”

mentioning

confidence: 99%

Surface plasmon resonance label‐free monitoring of antibody antigen interactions in real time

Kaušaitė

Dijk²,

Castrop³

et al. 2007

Biochem Molecular Bio Educ

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Detection of biologically active compounds is one of the most important topics in molecular biology and biochemistry. One of the most promising detection methods is based on the application of surface plasmon resonance for label-free detection of biologically active compounds. This method allows one to monitor binding events in real time without labeling. The system can therefore be used to determine both affinity and rate constants for interactions between various types of molecules. Here, we describe the application of a surface plasmon resonance biosensor for label-free investigation of the interaction between an immobilized antigen bovine serum albumin (BSA) and antibody rabbit anti-cow albumin IgG1 (anti-BSA). The formation of a self-assembled monolayer (SAM) over a gold surface is introduced into this laboratory training protocol as an effective immobilization method, which is very promising in biosensing systems based on detection of affinity interactions. In the next step, covalent attachment via artificially formed amide bonds is applied for the immobilization of proteins on the formed SAM surface. These experiments provide suitable experience for postgraduate students to help them understand immobilization of biologically active materials via SAMs, fundamentals of surface plasmon resonance biosensor applications, and determination of non-covalent biomolecular interactions. The experiment is designed for master and/or Ph.D. students. In some particular cases, this protocol might be adoptable for bachelor students that already have completed an extended biochemistry program that included a background in immunology.

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mentioning

confidence: 99%

Surface plasmon resonance label‐free monitoring of antibody antigen interactions in real time

Kaušaitė

Dijk²,

Castrop³

et al. 2007

Biochem Molecular Bio Educ

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“…Pancreatic proteases are divided into two major subgroups consisting of endopeptidases and exopeptidases. Endopeptidases including trypsin, chymotrypsin, and elastase are secreted into the duodenum as inactive precursors [ 46 ]. After being transformed into active forms, they catalyze the disruption of endopeptide bonds thus releasing relatively large oligopeptides.…”

Section: Protein Digestion In Animals Small Intestine As the Primarymentioning

confidence: 99%

Escherichia coli, an Intestinal Microorganism, as a Biosensor for Quantification of Amino Acid Bioavailability

Chalova

Sirsat

O’Bryan

et al. 2009

Sensors

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In animal diets optimal amino acid quantities and balance among amino acids is of great nutritional importance. Essential amino acid deficiencies have negative impacts on animal physiology, most often expressed in sub-optimal body weight gains. Over supplementation of diets with amino acids is costly and can increase the nitrogen emissions from animals. Although in vivo animal assays for quantification of amino acid bioavailability are well established, Escherichia coli-based bioassays are viable potential alternatives in terms of accuracy, cost, and time input. E. coli inhabits the gastrointestinal tract and although more abundant in colon, a relatively high titer of E. coli can also be isolated from the small intestine, where primary absorption of amino acids and peptides occur. After feed proteins are digested, liberated amino acids and small peptides are assimilated by both the small intestine and E. coli. The similar pattern of uptake is a necessary prerequisite to establish E. coli cells as accurate amino acid biosensors. In fact, amino acid transporters in both intestinal and E. coli cells are stereospecific, delivering only the respective biological l-forms. The presence of free amino- and carboxyl groups is critical for amino acid and dipeptide transport in both biological subjects. Di-, tri- and tetrapeptides can enter enterocytes; likewise only di-, tri- and tetrapeptides support E. coli growth. These similarities in addition to the well known bacterial genetics make E. coli an optimal bioassay microorganism for the assessment of nutritionally available amino acids in feeds.

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Chemical Digestion, Absorption, and Transport

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Gastrointestinal Physiology

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The zymogen‐enteropeptidase system: A practical approach to study the regulation of enzyme activity by proteolytic cleavage*

Cited by 5 publications

References 9 publications

Surface plasmon resonance label‐free monitoring of antibody antigen interactions in real time

Surface plasmon resonance label‐free monitoring of antibody antigen interactions in real time

Escherichia coli, an Intestinal Microorganism, as a Biosensor for Quantification of Amino Acid Bioavailability

Chemical Digestion, Absorption, and Transport

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