biochimia descriptiva incercdnd si atinga a: ai simplu si mai accesibil tuturor celor carte; 2 detalii pentru ca astfel materialul si fie sabilitatea s& fie cét mai mare; 2a de conexiuni cu lumea medicala real, ia fost posibil, eventualele modificari lular; +e apropierea limbajului biochimic de cel cartea mai atractiva; urs, acest material si poate fi folosit in 2a cht mai clara a proceselor metabolice, 2, de Ja nivelul organismului uman. fective au fost atinse si in acest fel cartea > vor parcurge si si fie de un real folos in Autorul 12 AMINO ACIDS AND PROTEINS Proteins are composed of several amino acids linked through peptide bonds. All proteins contain different sequences acids called the common amino acids. There are 20 common amin‘ the structure of proteins; other amino acids present in proteins de: the common amino acids. Most of the amino acids (except proline) are alpha (a) ami the amino group (NH2) is linked to the same carbon atom to v carboxyl group (COOH) is attached: RCH COOH ‘The general structure of amino acids NH, Classification of the amino acids (Common amino i A) Aliphatic amino acids: HH-COOH a, —cH-cooK Sen op coe NH, NH, NH, Giysine Alanine Vatne (Gly) (@) (Ala) (A) (van (v) ch, Sch—cH, —CH—CooH CH; —CH, —CH—CH—COOF cH, | NH CHy NH Leacine leans (Leu) (L) le) 13B) Hydroxy amino acids: : Gh—ch—cooH CH; —cH-CH—COOH OH NH OH NH, j Serine ‘Threonine 4 (Ser) (S) (Thr) (1) C) Sulphur containing amino acids: i ch cH —COOH CH, —CH, —CH—COOH SH NH, S—chH, NH, Cysteine Methionine 9) (et) a D) Acidic amino acids and their amid i Hooc—cH,—cH—COoH H,N— CO—CH, —CH—COOH NH, NH, Aspartic Acid nh (Asp) ®) aca Gn { HoOe—CHi—CH—cH—COOH HIN CO—CH,— CH —CH —CooH NH, NH, Glutamic Acid Glutamine Ct!) (Gin) (Q E) Basic amino acids: CH, Ch, —Cih —cH— COOH Ni, Gr, ~~: —G, —cH—COOH NH NH ! Arginine i Lysine NH E ys) 06) are) 4 Phet @he eae Pr.jcese dH, 1G co0H NH, ta —G4—cooH NH sine 1) = Sian Na NH NH Histidine is) a) F) Aromatic amino acids: i a € \ cr —ci—coon Ho cr, cH cool OF hh \/ | NH Phonylalanine ‘Tyrosine he) ) ayn) Oo I CH, CH —COOH Cyt. 8 Tryptophan en) W) G) Imino acids: N, coon B Proline (Pro) ®) 15Derived amino acids 1, Derived amino acids found in proteins These amino acids are found in some proteins and their structure derives from the structure of the common amino acids. After biosynthesis these proteins, some of the common amino acids are modified resulting these derived amino acids. Example of derived amino acids found in proteins: hydroxyproline, hydroxylysine, N-formylmethionine (fMet) HO—— L Ah CH, —CH—CH, —CH, —CH—COOH 7 COOH jane | N NH, OH NH, 4—Hydroxyprotine '5—Hydroxylysine (component of collagen) (component of collagen) Hh Cth 6H —COoH ionine (important in initiation of protein synthesis) 2. Derived amino acids not found in proteins ‘These amino acids are present in the cells as free amino acids (they fare not components of proteins), and they ate produced during the metabolism of amino acids. Examples of amino acids not found in proteins: homocysteine, homoserine, ornithine, citrulline, dihydroxyphenylalanine (DOPA), 7- aminobutyric acid (GABA), taurine, moniodotyrosine (MIT), diiodotyrosine (DIN triiodothyronine (15), tetraiodothyronine or thyroxine (Ts). oh —cH, SH Homocysteine (i methionine synt Ha CHa ie ae NH, th Che NH, B—Alanine pantothenic oh —CH; NH, 7-Aminot (the most 2 reurotrans Monoiodoty (precursorand their structure . After biosynthesis modified resulting ns: hydroxyproline, aH —COOoH NH, en) ve amino acids (they roduced during the eins: homocysteine, lanine (DOPA), 1 MIT), diiodotyrosine vxine (T,). ae eee SH NH, Homocysteine (intermediate in ‘methionine synthesis) CH, —CH, —CHl, —CH—COOH ia NH, C=O —— Citrattine Gavolved ia I turea synthesis) NH, Gt, —€Hy—COO# NH Alanine (component of pantothente acid) CH, —CH, —CH,— COOH NH) y—Aminobutyric acid (GABA) (the most abundant inhibitory neurotransmitter in the brain) on nen prcoom Monoiodotyrosine (MIT) (precursor of thyroid hormones) : (Ci, —CH; —FH COOH OH NH, Homoserine (intermediate in methionine, threonine, aspartic acid synthesis) CH, —CH, —-GH, —fH—ECOH NH, NH Ornithine (involved in urea synthesis) Ctl; CH SO4H NH, ‘Taurine (role in bile acids conjugation) OH on \po pico NH, Dihydroxyphenylalanine (DOPA) (volved in catecholamine syathesis) 1 On CH, —fH4—COOH 1 NH, Diiodotyrosine (DIT) (precursor of thyroid hormones)‘The isoelectric p — At ‘Triiodothyronine (13) (thyroid hormone) A) For 1 1 : ood op _— 1 1 NH, tH —! (Thyroxine) NHs Gh Properties of the amino acids lonization of the amino acids PH, ‘All the amino acids contain at least two ionisable groups: the a- carboxyl group (it is an acidic group — a group which can donate ‘H’) and the a-amino group (it is a basic group — a group which can accept H’). Depending on the pH of the medium, the amino acids can have a structure with maximum protons — the cationic form (with a positive charge), a structure with minimum protons ~ the anionic form (with a negative charge) or an isoelectric form-zwitterion (when the charge of the molecule is zero): R-CH-COOH 4H = R-CH-COO 2H” R-CH-COO" eS \ 3 | ‘NHS NERY Cation (+) “vterion Anion () (etructure with maximum (soelectrie form) (structure with minimum protons) o rotons) 18 OT STE!Chemical properties of the amino acids A) Reactions of the amino acids due to carboxyl group Al) Decarboxylation reaction: The amino acid is transformed into the corresponding amine with releasing of CO. nor GObH ——> RCH, —NH, + CO, NH, Amine Amino acid Some biologically important amines are produced in the body from amino acids: Histidine —> Histamine + CO, cH, —CH—COOH CH, —CH, | —, | + CO, NH Ny NH NH i NH Histidine Histamine ‘Tyrosine —> Tyramine + CO; ct, COOH th “<) NY + CO i Nit ‘Tyrosine : ‘Tyramine ‘Tryptophan —> Tryptamine + CO CH, —CH—COOH A CH, —CH, J | —f TT] 1 + co, " NH, S An, H H Tryptophan Tryptamine 22. Glutamic acid Hooc_cH, - Glutamic a A2) R radical of mor to form the ¢ proteins struct Hooc —cH,— Glutamic a HOOC—cH,— Aspartic ac A3) Es RH NH, Amino Acid The re synthesis.vacids | group 1 transformed into the the body from amino tt + CO, Nt, CH, Hy eo Hy CH, CH, Glutamic acid —> Gamma (y)-aminobutyric acid (GABA) + CO2 HOOC—CH, Ci, HCOOH + HO0C—CH, Ch “Hg NH, MES Glutamic acid Gamma (7) ~ aminobutyric acid (GABA) A2) Reaction with ammonia (NH): The -COOH group from the radical of monoamino dicarboxylic amino acids can combine with ammonia to form the corresponding amide (these amides are also components of proteins structure). HoOC—cH,—CH,—CHCOOH + NHy + H,N—CO—CH, CH, —CH_-COOH + 14,0 NH; Nis Glutamic acid Glutamine HOOC—cH-—CH—COOH + NH —s HiN—CO—CHi—CH—COOH + HO NH, NH, Aspartic acid Asparagine 3) Estherification reaction: a-GH-coG + ROB —>R-H_CO-0— 8 + H,0 NH, Alcohol NH, Amino Acid Esther ‘The reaction is important to protect the carboxyl group in peptide synthesis. 23|A4) Reaction with Lib R—CH—COOH) —HBH sR CH—CH, —OH } J NH NH, ‘Amino acid ‘Amino ~ aleobo! ‘The reaction is used for determination of the sequence of amino acids in proteins. B) Reactions of the amino acids due to amino group B1) Transamination reaction: The amino group of an amino is transferred to a keto acid forming a new amino acid and another keto acid. R-GH-COOH | Ry —-G-—COOH Trani COOH R,—CH-COOH + ——— * | NH, ° (Pyridoxal ° + NH ‘Amino acid Ketoacid —PBOSPHSE) Keto acid Amino acid ‘There are about twenty transaminases (or aminotransferases), but the two most important in our body (important in diagnostic) are: glutamate oxaloacetate transaminase (GOT) or aspartate aminotransferase (AST) and glutamate pyruvate transaminase (GPT) or alanine aminotransferase (ALT). The reaction catalyzed by GOT (AST) is the following: hooc-crcscr-coon + thode-ct:c-Coo -E Howe-cH;CH;e-COOH - MODE-CIti- COOH Ra, 0 6 Na The serum activity of GOT increases in: acute hepatitis, liver necrosis, myocardial infarction. 24 The re HOOC-CU:CHeC N conuranaien The s necrosis. B2) C acid is remov into the corre: In the glutamic acid Hooc: autas ccuuraan ‘The 1 which uses N. B3)R oe FDNBCH, —OH > the sequence of amino o group > group of an amino is and another keto acid. COOH Rr—CH-COOH + NH cid ‘Amino acid minotransferases), but the fiagnostic) are: glutamate tinotransferase (AST) and aminotransferase (ALT). te following: :Hy€-COOH + HO0C-CH-CH-COOH, 3 tn, Sac acm) caSPARTIC ACID) in: acute hepatitis, liver ‘The reaction catalyzed by GPT (ALT) is the following: HOOC-CHCHsCH-COOH + cH:C-COOH EE cUVCH-COOH + HOOC-CHsCHs¢-COOH Nu, 6 Gen NE é cura ymuvic Ac mee axsrosurtaeare (araae Aci) vnvarey (ektro atUTAMC ACI) The serum activity of GPT increases in: acute hepatitis, liver necrosis, B2) Oxidative deamination reaction: The amino group of an amino acid is removed as free ammonia (NHs) and the amino acid is transformed into the corresponding keto acid. R—CH—COOH 449 R—C—COOH + NH; | = il NH, 0 ‘Ammonia ‘Amino acid Keto acid In the body, the major oxidative deamination reaction is given by glutamic acid: ouvraMaTE HOOC-CH;CH-CALCOOH - |B HOOC-CH;CH;¢-COOH -(NE, o inn ouvrasare. |eXETOGLUTARATE (olvtaste Ac) (eKETO GLUTARIC ACID) The reaction is catalyzed by glutamate dehydrogenase, an enzyme which uses NAD* or NADP* as coenzyme. B3) Reaction with fluorodinitrobenzene (FDNB) (Sanger reagent): of \-+ + HN—cHcooH Ow SN | HE & R no, no; NH—c—cooH DNB Amino acid Dinitrophenyl derivative : 25The reaction is used to determine the N-terminal amino acid of a polypeptide. B4) Reaction with phenylisothiocyanate (PITC) (Edman reagent) I GWE + MEH coon GAH —F—N—cH- COOH, pire Amino 7 GH —H— eae s7NY7R } Phenylthiohydantoin derivative The reaction is used to identify the N-terminal amino acid of a polypeptide. BS) Reaction with dansyl chloride: Hyc-N—cHy HCN —CHy + H,N—CH—COOH +H I — Amino acid HN “ HCOOH Dansyl chloride Dansyl derivative ® The reaction is used to determine the amino acid from the N-terminal end of a polypeptide. B6) Reaction with formaldehyde: eco R—CH—COOH + CHO <—> | +H20 NH, N=CH, ‘Amino acid Formaldehyde Nitromethylene derivative 26 The: urine (Soren BT), ac NH, ‘Amino ine measuring tl BS) presence of MS c an ° Iierminal amino acid of a 1an reagent): <—e —CH—COOH 40 erminal amino acid of a +H HN —Fi—c00H iylderivative | ® o acid from the N-terminal CH—COOH 1, N=CH, romethylene ivative The reaction is used to determine the presence of the amino acids in ~ rine (Séreuisen method). BT) Reaction with HNOz: R—CH—COOH R—CH—COOH | + HONO ——~> | + N,+ H,0 NH, oH Amino acid Ihe reaction is used to determine the presence of an amino acid measuring the volume of resulted nitrogen (Van Slyke method). B8) Reaction with ninhydrin; Amino acids with ninhydrin, in the presence of heat, will form a complex, cae in pink, purple or blue. i ay SO Hi 6H coon Cy Lo 4b RACHOY Cp + Nhe Non fk me L Ko j i i i Myon Alcaye fi t a“ SO eat / i Loo oy © I 4 Coloured complex ©) Reactions of the amino acids due to carboxy: ss ass grOUpS C1) Peptide bond formation: HN—CH—COOH + Ha —CH—COOH + - + HiN—CH#—cooH fy Ry Ry Amino acid 1 Amino acid? Amino acid HN ci C0 -RHN—CH—CO oe HCOOH + (n-t)H,0 NA Polypeptide (Protein) a7