Amino Acids General Amino Acids: Building blocks for
Amino Acids General Amino Acids: Building blocks for peptides, proteins Some individually important (or converted to important molecules) Gly, Glu, Tyr neurotransmitters
Tyr parent/precursor for epinephrine (adrenaline) His stomach secretes HCl, symptoms for inflammation, colds. Essential (10) needed for normal health not synthesized by the body must be supplied by diet Complete (animal) vs. Incomplete (vegetable) protein
Complex proteins Multiple units Non-protein parts Metal ions
Primary Structure Primary Structure: #, kind, type, and sequence of AA Fredrick Sanger (1958 Nobel Prize) Beef Insulin Several years of work to sequence 51 AA Hydrolyzed proteins into smaller fragments to analyze
Fragment 1: Gly-Glu-Arg-Gly-Phe-PheGly-Phe-Phe-Tyr-Thr-Pro-Lys Fragment 2: Combined: Gly-Glu-Arg-Gly-Phe-Phe-Tyr-Thr-Pro-Lys Overlap Edman Degradation split AA at N-Terminal End
Secondary Structure Secondary Structure: Determined by H-bonds between AA-backbone -helix AA 4 residues apart, R-groups towards outside -pleated sheet AA far apart, R-groups face outwards
Tertiary Structure Tertiary Structure: Determined by interactions between R-groups H-bonds: -COOH and OH Ionic/Salt Bridges
Disulfide Bonds Hydrophobic (form core of protein) Hydrophilic (face outwards to interact with water) Quaternary Structure
Quaternary Structure: Multiple protein units Non-protein parts Metal ions Ex: Hemoglobin 4 subunits
Fe atoms Protein Structure Summary -Helix
-helix Structure:helix Structure: Secondary Determined by H-bonds between AA-backbone -helix AA 4 residues apart, R-groups towards outside -Pleated Sheet
-helix Structure:pleated sheet structure: Secondary Determined by H-bonds between AAbackbone -pleated sheet AA far apart, R-groups face outwards
H-bonds Secondary H-helix Structure:bonds: Between the C=O and NH of backbone Responsible for secondary structure Tertiary H-helix Structure:bonds:
Between the C=O and -NH or -OH of R-groups Responsible for tertiary structure Salt Bridges Ionic Bonds/Salt Bridges: Tertiary Structure
Between COO- and NH3+ groups Disulfide Bonds Disulfide bonds: Tertiary Structure Between -SH and SH groups
Mainly between Cys-Cys Hydrophobic Interactions Hydrophobic Interactions: Tertiary Structure
Between R groups (Alkane and Aromatic) Interior of proteins to avoid water Hydrophilic Interactions
Hydrophilic Interactions: Tertiary Structure Exterior of proteins to interact with water Polar groups (OH) Acidic groups (COOH) Basic groups (NH2)
Identify 2/3 Structure Protein Functions Protein Functions: Structural Support skin, connective tissue
Storage Fe in Liver Transport O2 in Hemoglobin Defense antibodies, venom Motion/Movement muscles Regulation blood/glucose/insulin Catalysis Enzymes (Ch. 30!)
Denaturation Denaturation: Loss of 3D conformation in a protein Disruption of 2/3/4 interactions Does NOT break 1 structure (hydrolysis) Loss of biological activity Causes of Denaturation
Xanthoproteic Test Xanthoproteic Test: Detects Benzene rings Yellow color
Phe, Try, Tyr Biuret Test Biuret Test: Detects tri-peptides (must have at least 2 peptide bonds)
Cu2SO4 Violet color Ninhydrin Test Ninhydrin Test:
General test for AA All AA blue Pro, hydroxyproline yellow Very sensitive 1 g (10g (10-6) Chromatography
Chromatography: separation technique for AA Difference in distribution between two phases Solubility Charge TLC (thin-layer) solid/liquid phase Solvent Front (rate solvent moves)
Differences in solubility cause AA to travel at different rates in the solvent Column chromatography (variation of above) Electrophoresis Electrophoresis: separation technique for AA
Charged particles separate in electric field (zwitterions) Separation based on Size friction (sieve) Charge electric field Types SDS masks charge/separate by mass/size Isoelectric Focusing AA separated by charge
2D separate on both. Fredrick Sanger Fredrick Sanger: Solved structure of beef insulin (1955) Nobel prize 1958
51 AA in two chains held together by disulfide bonds DFNB + N-terminal end + hydrolysis to solve structure Paper shredder
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