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Work Sheet 4 Digestion & Absorption of Proteins & Carbohydrates 1. Delineate the roles of gastric acid during digestion in the stomach. 1. Gastric HCl is responsible for the low pH <2 of gastric juice. 2. Gastric acid kills microorganisms and denatures dietary proteins preparing them for hydrolysis by proteases. 3. Gastric juices contain the acid stable proteases of the pepsin family, which produce large peptide fragments and some free amino acids. 4. Protein digestion at this stage is partial, as the amino acids and small proteins enter the duodenum, they trigger the release of cholectystokininpancreozymin (CCK-PZ) into the bloodstream. 2 List and distinguish the proteases involved in gastric, pancreatic, brush border surface and intracellular cleavage of peptide bonds; define their specificity in general terms. Gastric (Stomach) Digestion: Gastric juices contain the acid stable proteases of the pepsin family, which produce large peptide fragments and some free amino acids. Protein digestion at this stage is partial, as the amino acids and small proteins enter the duodenum, they trigger the release of cholectystokininpancreozymin (CCK-PZ) into the bloodstream. This release initiates the secretion of protease zymogens from the pancreas and release of enteropeptidase in the gut. Pancreatic Proteases: 1. The pancreatic juice is rich in the proenzymes of endopeptidase and the carboxypeptidases. 2. Enteropeptidase converts pancreatic trypsinogen to trypsin. 3. Trypsin autocatalytically activates more trypsinogen and other proenzymes, liberating chymotrypsin, elastase and the carboxypeptidases A and B. Digestion at the Brush Border (surface of intestinal epithelial cells): 1. Since pancreatic juice does not contain appreciable aminopeptidase activity, final digestion of di- and small peptides depends on brush border enzymes. 2. The surface of intestinal epithelial cells is rich in endopeptidases and aminopeptidases. 3. The end products of cell surface digestion are free amino acids and diand tripeptides. 3. Describe the absorption of proteins and the five transport systems. Absorption: 1. Following digestion, amino acids and small peptides are co-absorbed w/ sodium via group specific amino acids or peptide transport systems. 2. These processes are carrier mediated, they discriminate between natural L amino acids and D-amino acids, they require energy (from the Na+ gradient, Na-K ATPase) and physiologic temperatures. At least five brush border transport systems exist for: 1. neutral amino acids (uncharged aliphatic and aromatic) 2. basic amino acids (Lys, Arg, Cys, Cys-Cys) 3. acidic amino acids (Asp, Glu) 4. imino acids (Pro, Hydroxyproline) 5. di- and tripeptides 4. Explain the physiology of celiac sprue, cystinuria and Hartnup disease Clinical Correlates: 1. Hartnup Disease: Genetic defect in the neutral amino acid transporter. Symptoms: dermatitis due to tryptophan malabsorption ("niacin" flush) Consequences: not serious di- and tripeptide absorption supply minimal amounts of dietarily essential neutral amino acids. 2. Cystinuria: Precursor to kidney stones Symptoms: painful kidney stone formation due to malabsoprtion of cystine (two disulfide linked cysteines) 3. Sprue: Destruction and flattening of the intestinal villi resulting in generalized malabsorption. Causes: bacterial infection or gluten (contained in certain grains such as wheat and barley) sensitivity. 5. Compare and contrast the two polysaccharides of starch, amylose and amylopectin. SKIP 6. SKIP List the products of starch digestion by salivary and pancreatic -amylase. 7. Describe carbohydrate digestion and -glucosidases. General Information: 1. Carbohydrates provide a major component of the daily caloric requirement, ~40%. 2. Monosaccharides- do not need hydrolysis prior to absorption. Disaccharides- require brush border enzymes. Polysaccharides- require brush border enzymes, as well as, pancreatic amylase and salivary amylase for digestion. Enzymes Lactase Substrate Lactose Sucrase Sucrose Amylase Amylose Maltase Maltose Maltotriose -Limit Dextrins Isomaltose Isomaltase 8. Product Galactose Glucose Glucose Fructose Glucose Amylose (n-1) Glucose Maltose Glucose Distinguish between digestible and non-digestible carbohydrates. Undigested Carbohydrates: 1. Di-, oligo- and polysaccharides that are not hydrolyzed by -amylase and/or brush border enzymes cannot be absorbed. 2. These carbohydrates reach the lower tract of the intestine which contains bacteria. 3. The bacteria utilize many of the remaining carbohydrates, metabolizing them and producing by- products such as: hydrogen gas, methane and carbon dioxide. 9. Draw the products resulting from the sequential action of these peptidases: (1) Pepsin (2) Carboxypeptidase A (3) Elastase on the following recently ingested protein: H-Asp-Tyr-Gly-Glu-Phe-Lys-Asp-Val-OH Clearly indicate which fragments result from the action of each peptidase. The first one has been done for you. (6 pts) OH NH3 + CO2H CO2H O +H 3N C H C H 3C CO2H CH2 H N C H O H H N C O C H CH2 O H N C C H C H N C H O C O H N C H CH3 O H N C C H O H N C C H C (1) Pepsin: Endopeptidase, cleaves on the amino side of Tyr and Phe producing one free amino acid and two smaller peptides. OH NH3 + CO2H CH2 O +H 3N C H C H 3C CO2H CO2H H N C H O C H H N C H O C CH2 O N H C H C H N C H O C O H N C H C CH3 O O H N C H C H N Now draw the products resulting from digestion with Carboxypeptidase A on the above fragments. C H C O- O- Source Name Pancreas Trypsin Activator Enteropeptidase Trypsin Pancreas Chymotrypsin Trypsin Pancreas Elastase Trypsin Pancreas CP-A Trypsin Pancreas CP-B Trypsin Stomach Pepsin A H+, autoactivation Pepsin Trypsinogen Carboxy “R” Group vs. Specificity Amino C Arg,Lys Chymotrypsinogen Proelastase Pro-CP-A Pro-CP-B Pepsinogen A C C A A A Proenzyme Tyr,Phe,Trp Gly,Ser,Ala Val,Leu,Ile Arg,Lys Tyr,Phe