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INTRODUCTION TO,PROTEIN
Prepared byDr. Yogesh V. Ushir
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DEFINITION
Protein from the Greek proteios, meaning primary.
Proteins are the work horses of biological systems.–They play key roles in constructing and maintaining living cells
Our genes code for proteins
Proteins are polymers of amino acids
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SOURCES & QUALITY
Protein in food: different quantity & quality Nutritional value of protein: determined by
amino acid composition—Biological value (BV)
High biologicalvalueProtein that contains all essential amino acidsin sufficient proportions
Low biological valueProtein which is deficient (poor) in 1/moreessential amino acids
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ANIMAL & VEGETABLE PROTEIN
Animal proteinSources: e.g. meat, poultry, fish, eggs, milk, dairy products — Almost same proportion of each
essential amino acid as human protein —Complete protein: high BV
Vegetable proteinSources: e.g. vegetables, legumes, plants, grains, nuts, seeds— Deficient in 1/more essential amino acids—Incomplete protein: low BV — Deficient (poor) amino acid = limiting amino acid
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Animal protein = complete protein
Vegetable protein = incomplete protein
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MILK = HIGH QUALITY PROTEIN
Cow’s milk (and dairy products) = excellent source of high quality protein—All essential amino acids in proportions needed by the body
o Consists Whey protein (watery part of milk ) and casein (in milk salt of calcium )
Whey- liquid remaining after milk has been curdled and strained
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DIGESTION OF PROTEINS
Protein is hydrolysed (digested / cleaved) by particular protein-cleaving enzymes
Place of action: stomach & small intestine
Enzymes cleave protein in small units: —amino acids & short chain peptides
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ABSORPTION OF PROTEINS
Small units: absorbed by small finger-like projections (villi), lining the intestinal wall
Small peptides free amino acids
Free amino acids blood liver
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SYNTHESIS OF PROTEINS
Amino acids new (cell specific) protein = synthesis = continuing process
Body protein is continuously broken down (into amino acids) and re-synthesised (amino acids are re-built to protein)
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PROTEINS ARE:
Polypeptides + (cofactors, coenzymes, prosthetic groups, other modifications in animal)
Polypeptides are covalently linked a-amino acids
Cofactors are non-amino acid components e.g. metal ions like Zn2+in carboxypeptidase
Coenzymes are organic cofactors e.g. nucleotides in lactate dehydrogenase Prosthetic groups are covalently attached cofactors e.g. hemein myoglobin
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ROLES PLAYED BY PROTEINS INCLUDE
Enzymes (biologicalcatalysts) Hormones Storageproteins Transport proteins Structural proteins Protectiveproteins Contractile proteins Toxicproteins
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FUNCTION OF PROTEIN Major functional & structural component of all
cells in body required for growth, maintenance, renewal (repair) of all body cells
Functions as enzyme, transport carrier, hormone
—All enzymes, many hormones, blood transport molecules, hair, fingernails, etc. are proteins
Amino acids play a role as precursors (forerunner) for e.g. many enzymes, hormones, vitamins
Constituent of antibodies which protect against infection & disease
Supplies energy in particular circumstances (e.g. in malnourishment), —When fat & carbohydrate intake is
inadequate
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PROTEINS IN THE DIET
9 of the 20 amino acids must be obtained from the diet
These are referred to as the essential amino acids
Proteins are also the major source of nitrogen in the diet
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PROTEIN STRUCTURE
20 amino acids
Denatured
Quaternary
Tertiary
Secondary
Primary
Coded in DNA
Self assembly to a single (native) structure. Depends on primary structure and solution conditions
Common in foods. Many non-native forms depending on protein structure, solutionconditions (& history) and ingredient interactions
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Protein consists of amino acids
Amino acid (AA)= The basic organic compounds of proteins that are linked by peptide bonds (make up true protein).
Can be compared with a chain of beads---------etc
poly-peptide = protein Building unit =amino acid
A few amino acids linked together -------di-, tri-, or oligo-
peptides
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AMINO ACIDS FOLDED IN A 3-DIMENSIONAL STRUCTURE
Protein constituted by 5 elements in proportion; C = carbon - 50-55% H = hydrogen - 6-7% O = oxygen - 19-24% S = sulfur -0-4% N = nitrogen - 13-19% N= key element for
building tissues
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• Protein are polymers of a-amino acids• The amino acids used to make proteins are 2-amino-carboxylic acids•The α- (alpha) carbon is the carbon to which a functional group is attached.
PROTEIN STRUCTURE AND FUNCTION
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THERE ARE 20 STANDARD AMINO
ACIDS
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PROPERTIES OF AMINO ACIDS
structure and chemical functionality
Chirality
acid-base properties
capacity to polymerize
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AMINO ACIDS: 2 GROUPS
Essential amino acids—Must be supplied with food—Cannot be synthesized (made) by our body
Non essential amino acids
—Can be synthesized by our body
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ESSENTIAL (INDISPENSABLE) AMINO ACIDS
Isoleucine Leucine Lysine Methionine Phenylalanine Threonine Tryptophan Valine Histidine
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AMINO ACIDS: 5 GROUPS
Aliphatic chains: Gly, Ala, Val, Leu, and Ile
hydrophobicity
Hydroxyl or sulfur side chains: Ser, Thr, Cys, Met
Aromatic: Phe, Trp, Tyr
Basic: His, Lys, Arg
Acidic and their amides: Asp, Asn, Glu, Gln
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ALIPHATIC (ALKANE) AMINO ACIDS
Proline(pro, P) –cyclic “iminoacid”
Glycine(gly, G)–only non-chiralamino acid, not hydrophobic
Alanine(ala, A) –R-group = methyl-group
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Valine(Val, V) –Think V!
Leucine(Leu, L) –
Isoleucine(Ile, I) -2 chiralcarbons
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AROMATIC AMINO ACIDS All very hydophobic All contain aromatic group Absorb UV at 280 nm Phenylalanine(Phe, F) Tyrosine(Tyr,Y) –-OH ionizable(pKa= 10.5), H-
Bonding Tryptophan(Trp, W) –bicyclicindolering, H-
Bonding
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SULFUR CONTAINING AMINO ACIDS
Methionine (Met, M) –“start”amino acid, very hydrophobic
Cysteine (Cys,C) –sulfur in form of sulfhydroyl, important in disulfide linkages, weak acid, can form hydrogen bonds.
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ACIDIC AMINO ACIDS
Contain carboxyl groups (weaker acids than α-carboxyl-group)
Negatively charged at physiological pH, present as conjugate bases (therefore –ate not –ic acids)
Carboxyl groups function as nucleophiles in some enzymatic reactions
Aspartate–
Glutamate –
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BASIC AMINO ACIDS
Hydrophillic nitrogenous bases Positively charged at physiological pH Histidine–imidazole ring protonated/ionized, only
amino acid that functions as buffer in physio range. Lysine –di-amino acid, protonated at pH 7.0 Arginine-guianidinium ion always protonated, most
basic amino acid
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POLAR UNCHARGED AMINO ACIDS Serine(Ser, S) –looks like Ala w/ -OH
Threonine(Thr, T) –2 chiralcarbons
Asparagine(Asn, N) –amide of aspartic acid
Glutamine (Gln, Q) –amide of glutamicacid
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NONSTANDARD AMINO ACIDS
?
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These include the amino acid derivatives found in proteins;
Hydroxylysine, hydroxyproline -collagen Carboxyglutamate -blood-clotting proteins Pyroglutamate -bacteriorhodopsin Phosphorylatedamino acids -signaling device D-alanine, D-glutamic acid - bacterial cell wall
polypeptides Gamaaminobutyricacid (GABA)- neurotransmitter Homoserine -intermediate in amino
acid metabolism Ornithine - intermediate in
argininesynthesis
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FUNCTIONAL SIGNIFICANCE
Hydrophobic amino acids: encountered in the interior of proteins shielded from direct contact with water
Hydrophilic amino acids: generally found on the exterior of proteins as well as in the active centers of enzymes
Imidazole group: act as either proton donor or acceptor at physiological pH
–Reactive centers of enzymes Primary alcohol and thiol groups: act as
nucleophilesduring enzymatic catalysis–Disulfide bonds
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STEREOCHEMISTRY OF AMINO ACIDS
Amino acids are chiral (asymmetric)–Presence of an
asymmetric carbon atom
Natural proteins are made only from left-handed amino acids
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PEPTIDE BOND FORMATION0
9/0
7/2
01
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USE OF SYMBOLS IN REPRESENTING A PEPTIDES
In peptides the free –NH3+ is on the left while
free –COO – is on the right
Tripeptide –NH3
+ - glutamate-cystein-glycine-COO – - E - C – G one letter
symbols Glu- Cys – Gly three letter
symbols Glutamyl-cysteinyl-glycine peptide name
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PROPERTIES OF AMINO ACIDS PHYSICAL
Solubility Melting point Optical rotation Taste Acid base (ampholytes)
CHEMICAL Reaction due to –COOH grs.
Decarboxylation With ammonia
Reaction due to –NH2 grps. With ninhydrin Color reaction Transamination Oxidative deamination
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ACID-BASE PROPERTIES OF AMINO ACIDS
Amino acids are zwitterions:
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TITRATION OF AMINO ACIDS: ACID-BASE TITRATION
pK1: carboxylic acid = 2.4
pK2: amino group = 9.6
pI= (pK1+ pK2)/2
So, PI for leucine=6
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ABSORPTION SPECTRA OF AROMATIC AMINO ACIDS
Trp → λmax of 278nmTyr → λmax of 275nmPhe → λmax of 260nm
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DISULFIDE BRIDGE
- CH2 – SH + - CH2 – SH
- [O]
- CH2 – S– S - CH2 Two cysteine molecules under oxidizing conditions form the
Disulfide bridge Disulfide bonds between Cys residues stabilize the
structures of many proteins. Although Cys is a polar AA, the di-sulfide linked
residues (Cystine) are strongly hydrophobic.
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NINHYDRIN REACTION
– More common for amino acids
- Purple, blue or violet derivatives, 570nm
– Yellow for proline, 440 nm
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BONDS RELATING TO PROTEIN STRUCTURES
1. Strong bonds Peptide bond Disulphide bond
2. Weak bonds Hydrogen bonds- H-atoms between
nirogen & carbony oxygen Hydrophobic bonds- non-polar side
chains of neutral amino acids are closely associated with one another in proteins
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BIOLOGICALLY ACTIVE PEPTIDES Glutathione - 3peptides Thyrotropin releasin harmone - 3peptides Oxytocin - 9 amino acids Vasopressin (ADH) - 9 amino acids Angiotensins - 10 amino acids Methionine enkephalin - 5peptides Bradykinin - 9 amino acids Kallidin - 10 amino acids Aspatame - dipeptide Peptide antiibiotic- antibiotic such as gramicidine,
bacitracin, tyrocidin & actinomycin are peptide in nature
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PROTEINS
May be defined as the high molecular weight mixed polymer s of alpha-amino acids joined together with peptide linkage
-(CO – NH) –- The term protein is generally used for a
polypeptide containing more than (50 ?) 100 amino acids.
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CLASSIFICATION1. SIMPLE PROTEINS
a. Globular proteins: these are spherical or oval shape, soluble in water or other solvents and digestible
E.g.; albumin, globulin, glutelin, prolamines, histones, globins, protamines
b. Fibrous proteins: these are fibre like in shape, insoluble in water & resistance to digetion.
E.g.; collagens, elastins, keratins
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2. CONJUGATED PROTEINS a. Nucleoproteins:
nucleic acid (DNA & RNA) is the prosthetic group
E.g.; nucleohistones, nucleoprotamines
b. Glycoproteins: the prosthetic group is carbohydrates, which is less than 4 % of proteins. The term mucoproteins is used if > 4% of carbohydrates
E.g.; mucin, ovomucoid (egg white)
c. lipoproteins: the prosthetic group is lipid E.g.; serum lipoproteins, membrane LP
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2. CONJUGATED PROTEINS (CONTINUE…)
d. Phosphoproteins: phosphoric acid is the prosthetic group
E.g.; casein, viteline (egg yolk)
e. chromoproteins: prosthetic group is colored in nature
E.g.; Hb, cytochromes
f. metalloproteins: these protein contains metal ions, Fe, Zn, Cu, Mg
E.g.; ceruloproteins (Cu), carbonic anhydrase (Zn)
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3. DERIVED PROTEINS
a. Primary derived proteins: are the denatured or coagulated or first hydrolyzed product of proteins
E.g.; proteans (fibrin, fibrinogen), coagulated protein (cooked protein), metaprotein
b. Secondary derived proteins: are the progressive hydrolytic product of protein hydrolysis
E.g.; proteoses, peptones, polypeptides & peptides
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NUTRITONAL CLASSIFICATION
Complete proteins
Incomplete proteins
Already discuss earlier
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PROTEIN HYDROLYZING ENZYMES
Pepsin- in gastric juice
Trypsin-, chymotrypsin and carboxypeptidases- in pancreatic juice
Amino-peptidases, dipeptidases and polypeptidases-in intestinal juice
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STRUCTURE OF PROTEINS Primary- is the linear sequence of amino acids
forming the backbone of proteins (polypeptides)
Secondary- the spatial arrangement of proteins by twisting of the polypeptide chain
Tertiary- The three di-mentional structure of a protein
Quaternary- some of the proteins are composed of two or more polypeptides chains refereed to as subunits. The spatial arrangement of these subunits is known as Quaternary proteins
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TESTS FOR PROTEINS
a. Color reactions Biuret test Ninhydrin test
b. co-agulation reactions Heat coagulation
c. Ppt. reactions Gives gelatinous ppt with solid ammonium
sulphate
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ESTIMATION OF PROTEINS
Quantitative estimation of proteins of foods and other biological materials are performed by the following methods-
Kjeldhal method
Colorimetric method
Electrophoretic method
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Thank u… Thank u… meet u again meet u again in synthesisin synthesis
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