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DR/SA;;Y C.SUBA
Contents
1. 2. 3. 4. 5. Introduction Structures of Amino acids Metabolism of Amino acids Aminocidopathies Amino Acid Analysis
Introduction
i. Building blocks of proteins ii. Growth, repair and maintenance of cells
Peptide bond
Amino Acid
Chemical Properties
Ninhydrin Reaction
Ninhydrin( Triketohydrinthinhydrtae) is reduced to hydrindantin. The amino acid is dismuted to an aldehyde, ammnia, and carbon dioxide. The formation of colored chromogen is made by the reaction of Hydrindantin with ammonia and another molecule of ninhydrin .
Chemical Properties
XANTHOPROTEIC TEST- Using 65% nitric acid the aromatic
rings of amino acids like tyrosine and tryptophan are nitrated. The nitro derivate shows an intensely yellow color. Because nearly all proteins contain aromatics it is taken as a protein-test either. : Nitrated tyrosine (a) and tryptophane (b)
Chemical Properties
MILLONS TEST-This test, also known as Coles test, is answered by
compounds containing hydroxphenyl group. Tyrosine as shown in figure 7.43 both in free and bound forms react with Sodium nitrite under acid condition and is converted to nitrous acids. The end product is probably due to the formation of mercury phenolate with nitrated phenyl group
Chemical Properties
SAKAGUCHI TEST
Arginine, the only amino acid with a guanidium group that gives a red olour with napthol and Sodium hypobromitein the presence of an oxidizing agent such as Cl2 /Br2 .
Chemical Properties
SULFUR TEST By boiling with NaOH, S in the amino acid is converted into Na2S, which then precipitates as blackPbS with the addition of lead acetate.When proteins containing cysteine or cystine residues are boiled under strong alkali organic sulfur,it is converted to sulfide. Addition of lead acetate causes precipitation of insoluble black lead sulfide.
2. Monoaminodicarboxylic acids (acidic amino acids) Aspartic acid - aminosuccinic acid Glutamic acid - alpha amino glutaric acid
3. Aromatic Amino Acids (contain aromatic ring) Phenylalanine - alpha amino beta phenylpropionic acid Tyrosine - alpha amino beta parahydroxyphenyl propionic acid 4.Heterocyclic Amino Acids Tryptophan (also aromatic) - alpha amino beta indole propionic acid Histidine (also aromatic) - alpha amino beta imidazole propionic acid Proline (non-aromatic) alpha pyrolidine carboxylic acid Hydroxyproline (non-aromatic) - gamma hydroxyl alpha pyrrolidine carboxylic acid 5.Diamino Monocarboxylic acid (basic amino acids) Arginine - alpha amino lambda guanidinovaleric acid Lysine - alpha, epsilon diaminocaproic acid
These amino acids include alanine, asparagines, aspartate, cysteine, glutamate, glutamine, glycine, proline, serine and tyrosine.
Additional Information
i. Nutritionally essential amino acids is supplied by diet (e.g. histidine, lysine, etc.) ii. Some amino are produced in the body (e.g. cysteine, glutamine, etc.)
Nitrogen cycle
Processes involved in nitrogen cycle: Nitrogen Fixation. Molecular nitrogen (N2) is inert and does not react to form compounds. Substantial amounts of energy are needed to break it apart so that its atoms can combine with other atoms.
Nitrogen cycle
Nitrification. It involves the oxidation of the ammonium compounds in dead organic material into nitrites and nitrates by nitrifying bacteria.
Nitrosomonas bacteria can oxidize NH3 to nitrites (NO2) while Nitrobacter bacteria can oxidize the nitrites to nitrates (NO3)
.These bacteria live in the roots of legume plants such as peanuts, beans, and peas.
Nitrogen cycle
Denitrification. It is the process of reducing nitrates to nitrogen gas which replenish the nitrogen in the atmosphere
The carbon skeletons of the metabolites that enter the citric acid cycle will be completely oxidized into CO2 or diverted into gluconeogensis or ketogenesis.
Then glutamate is converted to glutamine which is catalyzed by glutamine synthetase: NH4+ + glutamate + ATP Pi glutamine + ADP+
Glutamate and Glutamine. Glutamate (glutamic acid) is synthesized from the transamination of -ketoglutarate which is catalyzed by glutamate dehydrogenase. Glutamine is formed when a second ammonium ion is incorporated into glutamate by the action of the enzyme glutamine synthetase:
Ketogenic
Lysine leucine
Glycine Serine Valine Histidine Arginine Cysteine Proline Alanine Glutamate Glutamine Aspartate Asparagine Methionine
Thyroxine is a hormone synthesized from tyrosine which is secreted by the thyroid gland that regulate metabolism.
Aminoacidopathies
Class of inherited errors of metabolism Enzymes defects that inhibits the bodys ability to metabolize certain amino acids
i. ii. iii. iv. v. PKU Tyrosinemia Alkaptonuria MSUD Isovaleric Acidemia
vi. vii. viii. ix.
Aminoacidopathies
Aminoacidopathies
i. Phenylketonuria
Absence of phenylalanine hydroxylase (PAH)
PAH
1.
2.
Laboratory Tests
Guthrie test
Microfluorometric assay
Aminoacidopathies
i. Phenylketonuria
1.
Laboratory Tests
Guthrie test Semi quantitative bacterial inhibition assay Uses phenylalanine to facilitate bacterial growth (B. subtilis and -2-thienylalanine).
Aminoacidopathies
i. Phenylketonuria
2.
Laboratory Tests
Microfluorometric assay Direct quantitative measurement Phenylalanine ninhydrin copper fluorescence in the presence of dipeptide (360-530 nm)
Aminoacidopathies
ii. Type II Tyrosinemia
Tyrosine aminotransferase
Aminoacidopathies
v. Alkaptonuria
Lack of homogentisate oxidase
Phenylalanine hydroxylase
Phenylketonuria
Tyrosine aminotransferase
Tyrosinemia II
Tyrosinemia III
4-hydroxyphenylpyruvate dioxygenase
Alkaptonuria
Homogentisate oxidase
Fumarylacetoacetate hydrolase
Tyrosinemia I
Aminoacidopathies
vi. Maple Syrup Urine Disease
branched-chain -ketoacid decarboxylase
Aminoacidopathies
vi. Maple Syrup Urine Disease
1.
Laboratory Tests
Modified Guthrie test uses branched chain -ketoacid to facilitate bacterial growth (containing B. subtilis and 4-azaleucine).
Aminoacidopathies
vii. Isovaleric Acidemia
Deficiency of isovaleryl-CoA dehydrogenase
Aminoacidopathies
viii.Homocystinuria
Lack of cystathionine - synthetase
Aminoacidopathies
viii.Homocystinuria
1.
Laboratory Tests
Modified Guthrie test Uses methionine to facilitate bacterial growth (containing B. subtilis and L-methionine sulfoximime).
Aminoacidopathies
ix. Citrullinemia
i.
Type I citrullinemia
Lack of arginonosuccinic acid synthetase (ASAS)
Aminoacidopathies
ix. Citrullinemia
i.
Type II citrullinemia
Mutation of the gene that encodes for protein citrin
Aminoacidopathies
x. Argininosuccinic aciduria
Lack of argininosuccinic acid lyase (ASL)
Aminoacidopathies
ix. Citrullinemia x. Argininosuccinic aciduria
Aminoacidopathies
xi. Cystinuria
Defect in amino acid transport system by inadequate reabsorption of cystine in the kidneys
Aminoacidopathies
Class of inherited errors of metabolism Enzymes defects that inhibits the bodys ability to metabolize certain amino acids
i. ii. iii. iv. v. PKU Tyrosinemia Alkaptonuria MSUD Isovaleric Acidemia
vi. vii. viii. ix.
Disease
PKU
Enzyme deficiency
Phenylalanine hyroxylase Tyrosine aminotranferase 4-Hydroxyphenylpyruvate oxidase
Tyrosinemia I
Tyrosinemia II Tyrosinemia III
Alkaptonuria
MSUD
Homogentisate oxidase
Branched-chain -ketoacid decarboxylase
Isovaleric acidemia Isovaleryl-CoA dehydrogenase Homocystinuria Citrullinemia Cystathionine- synthetase Arginosuccinic acid synthetase Argininosuccinic acid lyase Defective AA transport system
Arginosuccinic aciduria
Cystinuria
Aminoacidopathies
Diagnosis
i. ii. iii. iv. v. 6-8 hours fasting Collected in heparin tube with the plasma Deproteinization performed within 30 minutes Screening by TLC stained with ninhydrin (blue) Separated and quantitated by Ion exchange chromatography and HPLC reversed-phase system or capillary electrophoresis