Structure and properties of proteins. Vladimíra Kvasnicová



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Structure and properties of proteins Vladimíra Kvasnicová

Chemical nature of proteins biopolymers of amino acids macromolecules (M r > 10 000)

Classification of proteins 1) by localization in an organism intra- / extracellular 2) by function structural / biological active 3) by shape globular / fibrous 4) by chemical composition simple / complex (conjugated) proteins

conjugated proteins contein polypeptide chain + prosthetic group: glycoproteins + saccharide metalloproteins hemoproteins phosphoproteins nucleoproteins (lipoproteins) + metal ion + heme + phosphoric acid + nucleic acid + lipids

Proteins are synthesized from L-α-amino acids 21 proteinogenic AAs other AAs are non-proteinogenic The figure was adopted from: J.Koolman, K.H.Röhm / Color Atlas of Biochemistry, 2 nd edition, Thieme 2005

Side chains of AAs determine final properties of proteins Isoelectric point (pi) = ph value at which the net charge of a compound is zero pi = (pk COOH + pk NH3+ ) / 2 Solutions of AAs belong among ampholytes AMPHION (= amphoteric electrolytes)

pk a Henderson-Hasselbalch equation: acid base + H + (for example: R-COOH R-COO - + H + ) If ph of a solution (e.g. cytosol) is higher than pk a (i.e. more basic) the structure of base predominates: acid base + H + The figure was adopted from: J.Koolman, K.H.Röhm / Color Atlas of Biochemistry, 2 nd edition, Thieme 2005

The figure was adopted from: J.Koolman, K.H.Röhm / Color Atlas of Biochemistry, 2 nd edition, Thieme 2005

Functional groups of amino acids -COOH -COO - + H + -COOH -CO- -NH 2 + H + -NH + 3 -CONH 2 (not basic) -OH (not acidic in H 2 O) -SH (not acidic in H 2 O) -SeH -S- -HN-C(=NH)-NH 2 carboxyl group anion carboxyl group acyl amino group cation amide group alcohol group thiol group selenol group sulfide group guanidine group

Important reactions of AAs 1) dissociation bases 2) decarboxylation biogenic amines 3) transamination 2-oxoacids 4) deamination 2-oxoacids 5) formation of peptide bonds peptides or proteins H H peptide bond

Amino acids in proteins L-α-aminocarboxylic acids 21 proteinogenic AAs other AAs are formed by a posttranslational modification

Modified amino acids The figure was adopted from: Devlin, T. M. (editor): Textbook of Biochemistry with Clinical Correlations, 4th ed. Wiley-Liss, Inc., New York, 1997. ISBN 0-471-15451-2

Important groups of proteinogenic amino acids branched chain AAs: valine, leucine, isoleucine aromatic AAs: phenylalanine, tyrosine, tryptophane, histidine sulfur-containing AAs: methionine, cysteine alcohol group containing AAs: serine, threonine, tyrosine basic AAs: lysine, arginine, histidine acidic AAs: aspartic acid (aspartate), glutamic acid (glutamate) amide group containing: asparagine, glutamine not optically active: glycine rare amino acid: selenocysteine abundant in blood: glutamine, alanine

Essential amino acids essential in a diet, they are not synthetized in a human body 1) branched chain AAs (Val, Leu, Ile) 2) aromatic AAs (Phe, Trp) 3) basic AAs (Lys, Arg, His) 4) secondary-oh (Thr), sulfide group (Met)

Peptides and proteins contain 2 or more AAs bound by peptide bond(s) common names are used systematic names: AA 1 -yl-aa 2 -yl-aa 3 oligopeptides: polypeptides: 2 10 AA > 10 AA proteins: polypeptides of M r > 10 000

border: polypeptide /protein is not sharp (~ 50 AAs) AAs are bound by peptide bonds the order of AAs in a chain (= primary structure) is given by a genetic information the order of AAs is reported from N- to C- terminal

Description of structure of proteins the macromolecule contains various AAs, in an exactly defined order and quantity spacial arrangement and biological function are DEPENDENT on the amino acid composition native protein biological active conformation

Side chains of AAs influence a final structure of proteins: polar side chain nonpolar side chain final conformation of the protein in water unfolded protein

the peptide chain has a special spatial arrangement: only some proteins are composed of subunits (= quaternary structure)

Bonds found in proteins 1) covalent peptide bond disulfide bond -CO-NH- -S-S- 2) noncovalent interactions hydrogen bonds -H... O- -H... N- hydrophobic interactions nonpolar side chains ionic interactions -COO - / + H 3 N-

Primary structure of proteins = order of amino acids read: from N-to C- end it is coded on a genetic level stabilization: peptide bonds

Secondary structure of proteins = spatial arrangement of the polypeptide chain given by rotation of the planar peptide bonds around α-carbons stabilization: hydrogen bonds between CO- and -NHof the peptide bonds α-helix β-pleated sheet β-turn real proteins: different parts of the polypeptide chain exist in various secondary structures

Helical structure (helix) various types of the spiral: different steepness, direction of rotation, number of AAs per turn peptide bond planes are parallel to the axis of the helix with R- perpendicular to it H-bonds are formed between AAs found above and below themselves the most common: α-helix (right-handed) collagen helix (left-handed, steeper)

β-pleated sheet (β-structure) direction of parts of the polypeptide chain is either parallel or antiparallel N C N C N C C N R- are placed above or below the plane of the sheet H-bonds are formed between peptide bonds of the neighboring parts of the polypeptide chain it brings strength to proteins

β-bend (reverse or β-turn) reverse the direction of a polypeptide chain, helping it form a compact, globular shape often connect successive strands of antiparallel sheets Nonrepetitive secondary structure loop or coil conformation not random but less regular structure than α- or β- one half of a protein molecule exist in it

Tertiary structure of proteins = spatial arrangement of the secondary structures (folding of domains) stabilization: between side chains of AAs 1) hydrogen bonds 2) ionic (electrostatic) interactions 3) hydrophobic interactions 4) disulfide bonds

tertiary structure secondary structures

α-helix β-sheet motif: barrel

Classification of proteins according to their tertiary structure 1) globular proteins (spheroproteins) spheroidal shape both secondary structures are abundant 2) fibrous proteins (scleroproteins) rod-like shape one secondary structure predominates e.g. α-keratin, collagen

Quaternary structure of proteins = oligomeric structure of a protein (2 or more subunits ~ monomers) i.e. the structure is found only in proteins composed from 2 or more chains (subunits) stabilization: noncovalent interactions the proteins have an allosteric effect

SUMMARY of protein structure description

properties and functions of proteins are related to their spatial arrangement IT DEPENDS ON AMINO ACIDS COMPOSITION funkční domény

Physicochemical properties water solubility depends on the structure proteins form colloidal solutions (viscosity, sedimentation, light dispersion) colloidal-osmotic pressure = onkotic pressure proteins can be salting-out of the solution (~ water sheet removing)

proteins can be denaturated heat, whipping, shaking, radiation strong ph changes, salt of heavy metals, organic solvents, detergents

proteins strongly absorb UV radiation

proteins are ampholytes -COOH -COO - + H + -NH 2 + H + -NH 3 + under physiological ph proteins are negatively charged ANIONS