BIOCHEM 321 FINAL EXAM CHEATSHEET
IgG: 2 heavy chains, 2 light chains held together by disulfide bonds and noncovalent interactions, C region bound to macrophage.
WESTERN BLOT: get lysate with protein of interest, run in SDS gel to separate by size, transfer proteins to a membrane, pass sol with a specific antibody on it
MYOSIN= left handed coil with large globular heads. head=binding site for nucleotide+actin. When ATP binds to myosin pocket, myosin dissociates from actin. Then ATP-> ADP +P and
myosin rebinds
ALPHA HELICES BETA SHEETS/STRANDS PROTEIN PURIFICATION A KERATIN=
rise=5.4 amperes, pitch=1.5 amperes rise=3.5 amperes, pitch=7 amperes Extraction → Differential Centrifugation → Salting Out → coiled coil of alpha
h bonds parallel h bonds ⟂ to the direction of beta strand Dialysis → Chromatography. helices, 3.6 AA/turn
n bonds to carboxyl group: AA-4 length of beta strand=#AA*3.5 amperes ● Salting out=adding salt to decrease solubility COLLAGEN=
carboxyl bonds to n: AA+4 antiparallel=more extended (3.5 (outcompete H2O interactions) and precipitate polypeptide chains that
#h bonds in alpha helice= #AA-4 amperes vs 3.2) w/ stronger hydrogen out proteins. are twisted around each
length of alpha helix= #AA * 1.5 amperes bonding (perpendicular instead of ● Size exclusion chromatography=LARGER other, left handed coil, 3
turns of alpha helix= #AA/3.6 or L/5.4 amp diagonal) PROTEINS ELUTE FIRST (opposite of gel AA/turn, rich in proline
bad for alpha helices: pro, gly, charged R group, Adjacent residues are furthest apart, electrophoresis). and gly
positive things near n terminal, large groups like look for 1 side polar ● SDS page=SMALLER PROTEINS ELUTE SILK FIBROIN=beta
trp, tryp, phe FIRST, separation based on size only sheets,spiders, rich in
**AA on same face: AA 1 and AA -5 ala and gly
**look for hydrophobic faces, MALQK
TYPES OF ENZYMES/CATALYSIS DENATURATION
Types of enzymes: OTHILL. Hydrolase radiation, changes in
cleaves bonds by adding H2O, lyase temp/ph, adding
cleaves/forms bonds w/ addition and reagents like urea and
elimination reactions (adds double bonds b mercaptoethanol
and rings). Ligases use ATP.
Types of catalysis: acid-base, metal ion,
covalent, electrostatic
HEMOGLOBIN
2 alpha, 2 beta subunits
When O2 binds, Fe shrinks, more
planar conformation. Fe2+=good,
more BPG at HA
CO=nonsigmoidal curve
Histidine does not stabilize CO
Co2 binds to Nterminus
Each chain has: multiple a helices,
1 pocket, a hydrophobic core
CATALYTIC EFFICIENCY
kcat/km. Want 10^8 or 10^9
inverse molarity* inverse seconds
tissues=high co2, low pH (many H+ ions) to release O2 (histidine=protonated) lungs= low co2, high pH to bind O2
kcat*[ET]=vmax
IgG: 2 heavy chains, 2 light chains held together by disulfide bonds and noncovalent interactions, C region bound to macrophage.
WESTERN BLOT: get lysate with protein of interest, run in SDS gel to separate by size, transfer proteins to a membrane, pass sol with a specific antibody on it
MYOSIN= left handed coil with large globular heads. head=binding site for nucleotide+actin. When ATP binds to myosin pocket, myosin dissociates from actin. Then ATP-> ADP +P and
myosin rebinds
ALPHA HELICES BETA SHEETS/STRANDS PROTEIN PURIFICATION A KERATIN=
rise=5.4 amperes, pitch=1.5 amperes rise=3.5 amperes, pitch=7 amperes Extraction → Differential Centrifugation → Salting Out → coiled coil of alpha
h bonds parallel h bonds ⟂ to the direction of beta strand Dialysis → Chromatography. helices, 3.6 AA/turn
n bonds to carboxyl group: AA-4 length of beta strand=#AA*3.5 amperes ● Salting out=adding salt to decrease solubility COLLAGEN=
carboxyl bonds to n: AA+4 antiparallel=more extended (3.5 (outcompete H2O interactions) and precipitate polypeptide chains that
#h bonds in alpha helice= #AA-4 amperes vs 3.2) w/ stronger hydrogen out proteins. are twisted around each
length of alpha helix= #AA * 1.5 amperes bonding (perpendicular instead of ● Size exclusion chromatography=LARGER other, left handed coil, 3
turns of alpha helix= #AA/3.6 or L/5.4 amp diagonal) PROTEINS ELUTE FIRST (opposite of gel AA/turn, rich in proline
bad for alpha helices: pro, gly, charged R group, Adjacent residues are furthest apart, electrophoresis). and gly
positive things near n terminal, large groups like look for 1 side polar ● SDS page=SMALLER PROTEINS ELUTE SILK FIBROIN=beta
trp, tryp, phe FIRST, separation based on size only sheets,spiders, rich in
**AA on same face: AA 1 and AA -5 ala and gly
**look for hydrophobic faces, MALQK
TYPES OF ENZYMES/CATALYSIS DENATURATION
Types of enzymes: OTHILL. Hydrolase radiation, changes in
cleaves bonds by adding H2O, lyase temp/ph, adding
cleaves/forms bonds w/ addition and reagents like urea and
elimination reactions (adds double bonds b mercaptoethanol
and rings). Ligases use ATP.
Types of catalysis: acid-base, metal ion,
covalent, electrostatic
HEMOGLOBIN
2 alpha, 2 beta subunits
When O2 binds, Fe shrinks, more
planar conformation. Fe2+=good,
more BPG at HA
CO=nonsigmoidal curve
Histidine does not stabilize CO
Co2 binds to Nterminus
Each chain has: multiple a helices,
1 pocket, a hydrophobic core
CATALYTIC EFFICIENCY
kcat/km. Want 10^8 or 10^9
inverse molarity* inverse seconds
tissues=high co2, low pH (many H+ ions) to release O2 (histidine=protonated) lungs= low co2, high pH to bind O2
kcat*[ET]=vmax
