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Proleins: Chem[siry And Living things are made up of many different chemical molecules. One important group of chemical molecules is proteins. Proteins make up the bulk of all solid material within your body and the bodies of other animals, your muscle, skin, hair, and inside organs are largely protein. Proteins are essential for body growth and repair. They also make up some hormones which function in chemical control in the bod~r. In th~s L~vestigation, you will (a) learn how to recognize molecular formulas for small molecules called amino acids. [b) use models of different amino a~ids to construct a protein molecule. (c) use chemical tests to determine Lf protein is or is not present in different substances. Materials paper models SCiSSOrS dropper glass marking pencil or labels test tubes test tube rack Ior tin can} nitric acid fingernail clippings egg white ihard-bofled} absorbent cotton dog hair {white] Procedure FIGURE 7-1 Part Ao Models of Protein I. Amino Acids, Building Blocks of Protein ....... a~I~-~eins are complex moleanles made .up of smaller molecules called amino acids. There are about twenty different amirio acids fomad in nature. The element nitrogen IN} is present in all anxino acids. Examine the structural forrdulas of the four representative amino acids shown in Figare 7-1. HHo HHo I ! tt H--N--C--C--O--H H glyclne alanMe H H 0 HHo II 1o Name.the four elements pre~ent in th~g-amix~o H--O~C--H I acids. H valise threonlne 27 What is the molecular formula for the amino acid la) glycine? C__H__O__N__ Use the paper models given to you by your teacher to complete this section. (c) valine? ® Cut out the four amino acid models. CAUTION: Always be extremely careful w~th scissors. Cut along the solid Bnes only. Attempt to join the amino acids. Id) threonine? 8. Can the amino acid models easily join to form a (b) alar,dme? How do the molecular formulas for all the amino acids differ? Note the upper fight comer of each amino acid. These ends have a special arrangement of carbon, oxygen, and hydrogen atoms. This end arrangement is called a carboxyl group and looks like this: protein molecule? ® loin the molecules by removing as many --OH groups and --H groups as needed from the amino acids. All four amino acid molecules can be joined in this manner to form a protein. Join them in the order valine--threonine--alanine--glycine. ® loin the leftover --OH and --H ends. 9. What chemical substance is formed when the --OH’s and --H’s are ioined? 10. How many molecules of water are formed when four amino acids ioin? 4. Circle the carboxy! group on each structural formula in Figure 7-1. Note the upper left comer of each amino acid. These ends have a special 11. What chemical compound is formed when the arrangement of nitrogen and hydrogen atoms. four amino acids are ioined? The end arrangement is called an amino group and looks like this: o 12. Describe the difference between an amino acid molecule and a protein molecule. 5. Use dashed lines to circle the amino groups on the structural formulas in Figure 7-!. There are thousands of di~{erent proteins in ft. In lab 6, you studied caxbohydrates. livIng organisms. What makes each protein differ(a) Do carbohydrates have carboxyl groups.~ ent is the order, number, kind, and arrangement in space of amino acids joined. You only assembled four amino acids into a protein using a specific (b) Do carbohydrates have amino groups? order. 7. HOW does the number of hydrogen atoms compare to the number of oxygen atoms in eaeh 13. Construct two proteins different ~rom the one you made above. List the order of amino acids amino acid? ) II. Combining Amino Acids to Form Protein Amino acids are not protein molecules. They re only the "building blocks" of protein. Several uino acids must be chemically joined in a chain form a protein molecule. We can show how no acids join by using models. (b) Part B. Identification of Proteins o Number five clean test tubes 1 to 5. Place them in a test tube rack. Using Figure 7-2 as a guide, add the ~ollowing substances to each test tube: tube l--fingernail clippings tube 2--egg white, hard-boiled tube 3--absorbent cotton tube 4--dog hMr, white tube S--cream cheese e Add 5 drops of nitric acid to each test tube. CAUTION: Nitric acid is harmful to skin and clothdng. Rinse w~th water ff spillage occars. Call you~ teacher. The test used to identi~ protein is technically called the xanthoproteic test. A substance conraining protein will turn yellow when nitric acid is added to it. No color change to yellow indicates that the substance being tested has no protein. ® Wait several minutes. Thin record the color of the items placed in each robe in Table 7-1. e On the basis of the xanthoproteic test, indicate in.the last column of the table i~ the substances tested do or do not contain protein. FIGURE 7-2 fingernail egg clipping~ white cotton white dog hair cream cheese PROTEIN TEST TABLE 7ol. TESTING SUBSTANCES TO DETERMINE ~F PRoTEiNS ARE PRESENT SUBSTANCE Fingernail Egg white, C~tton Dog hair Cream cheese COLOR CHANGE DUE TO NITRIC ACID SUBSTANCE TESTED iS A PROTEIN (ANSW£R YES OR NO) Part B. Tests for Lipids Brown Paper Test for Liplds 1. Place a drop of water on a small piece of brown paper. Place a drop of oil on the same piece of paper. Allow the paper to dry for a few minutes. 2. Hold the piece of paper up to the light. If a semi-transparent (translucent) spot is evident, the sample contains lipids. Record the appearance of the two spots in Table 3. Solubility Test for Lipids 1. Label two test tubes 1 and 2. 2. Using separate droppers, add 20 drops of 95% ethanol to test tube I and 20 drops of water to test tube 2. 3- Add 5 drops of oil to test tubes 1 and 2 and stopper each tube. 4. Shake each tube well, let settle, and record in Table 4 whether the oil is soluble in either solvent. Lipids are soluble only in nonpolar solvents because lipids, themselves, are nonpolar. Dispose of the contents of the test tubes according to your teacher’s directions. Gently use a test-tube brush and soapy water to clean the two test tubes and rinse with clean water. Par~ C. Tests for Proteins 1o Label three test tubes 1, 2, and 3. 2. Using separate droppers, add 30 drops of 2% gelatin to test tube 1, 30 drops of glucose solution to test tube 2, and 30 drops of water to test tube 3. Record the color of each tube’s contents in Table S. S. Add 10 drops of biuret reagent to’each test tube. CAUTION: Biuret reagent is extremely caustic to the skin and clothing. If biuret reagent is spilled, rinse with water and call " your teacher immediately. When biuret reagent is mixed with a protein, it will produce a lavender to violet color. Record in Table 5 the color of each tube’s contents atter adding biuret reagent. Discard the contents of the test tubes according to your teacher’s directions. Gently use a test-tube brush and soapy water to clean the test tubes and rinse with clean water. Fill in the last column of all live tables with the correct interpretation of the test results. Table 3. ffi Brown Paper Test for Lipids Translucent on brown paper? Liplds present(+/-) Substance Water Oil Table 4. Solubility Test for L~pids Substance Dissolves? Lipids present (+/-) Oil in ethanol Oil in water Table Test for Proteins Test tube Substance 1 Gelatin Color at start Color after adding bluret reagent Protein present Glneose Water 6. What is used to test for the presence of protein? 7. How can you tell by using this test that a substance contains protein? 8. Biuret reagent will turn the skin brownish-purple. Explain why this occurs. 9. a. When greasy food is spilled on clothing, why is it difficult to clean with water alone? - b. What would be better than plain water for removing a greasy food stain? Why? Analysis Use your results from Part A to answer questions 1-8. 1. Name four amino acids. 2. (a) How many amino ands are there? (b) How are amino acids used by living things? 3. List several of your body parts that are protein. 4. Name the four chemical elements p~esent in the amino acids studied (and in all amino acids). 5. Name the two special end groups present in amino acids. 6. What element is present in protein (amino acids) that was not present in carbohydrates (Laboratory Investigation 6)? 7. Explain how a protein molecule is formed in living organisms. 8. Explain how one protein differs from another protein. .... Use your Iesults from Part t3 to answer questions 9-13. 9. Describe how to tel1 if ~ substance is a protein by using the xanthoproteic test. __ 10. (ai List those substances tested that were protein. (b) List those substances tested that were not protein. 11. Using what you have learned about proteins, decide which of the fo!lowing substance,.s are protein. Place a checkmark on the line next to each substance that is protein. la) hamburger (ei liver. (b) chicken (f) human hair (el peanut oil (gl stomach Id) maple syrup {h) 207 amino acids joined 12. In Latin, the word "xantho" meana yellow, and "’proteic" means protein. Why is "xanthoproteic" a meaningfial word to use when describing the chemical test used for identifying a protein.~ 3O