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what is sciene
the process of gaining knowledge and understanding about the natural world
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practical side of scientific approach
- experimental reproducibility
- honesty
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fundamental assumption of science
forces and phenomena existing within the universe can be understood by applying a process of inquiry-the scientific method
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man in history who introduced reproducibility
alhaven
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possible phases of evolution of life
- prebiotic chemistry
- self-replicating molecules
- an RNA world (catalysis and information)
- breakthrough organism (package with crude metabolism)
- progenote (L.U.C.A.)
- biological evolution
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timeline of evolution
- earth formation 4.6 bil
- oldest fossil 3.7 bil
- evidence of O2(photosynthesis) 2.5 bil
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palimpset
- something that's been written over and over again
- ex: cofactors or enzymes
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hydrophobic interaction of water and oil
in the presence of water oil will build up and coagulate and pack tightly to block out the polar water molecules
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how to find pH from proton concentration
pH=-log[H+]
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henderson hassel-balch equation
pH=pK-log[acid]/[base]
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the stronger the acid...
the stronger the pKa
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non-polar aliphatic R groups
glycine, alanine, valine, leucine, isoleucine, methionine, proline
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Polar, uncharged R groups
serine, threonine, cysteine, asparagine, glutamine
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aromatic R groups
phenylalanine, tyrosine, tryptophan
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positively charged R groups
lysine, arginine, histidine(aromatic)
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negatively charged r groups
aspartate, glutamate
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configuration
molecule can be inter converted only by breaking and reforming one or more covalent bonds
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conformation
can be interconverted without breaking a covalent bond ex: rotation around sigma bond
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UV absorption amino acids
tryptophan, tyrosine, phenylalanine
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peptide bond formation in vitro
- attached to resin
- removal of protecting group
- activation of next amino acid
- peptide bond formation
- removal of polypeptide from resin
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how do peptides form invitro
carboxyl to N-temrinus
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function of proteins
- catalysis
- transport
- nutrient/storage
- motion
- structure
- defense
- regulation
- misc.
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which amino acid forms disulfide bonds
2 cysteines forms cystine
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avg. UV absorption for protein
OD280 0.55 for a 1 mg/mL solution
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ways to detect protein
UV absorption and colorimetric assays
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international enzyme unit
1 micromol=10-6 amount of enzyme that wil convert 1 micromol of product in one minute at 30 C
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units for protein activity
units/mL
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units for protein concentration
mg/mL
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units for protein specific activity
enzyme units/ mg of protein
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protein purification procedure
- break open cells
- fractionate
- test for purity
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precipitate nucleic acids with
- positively charged polymer
- polyethylene glycol
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precipitate protein with
- salt(NH4SO4) least harmful
- pH extremes
- heat
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types of chromatography
- ion-exchange
- size exclusion
- affinity chromatography
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