Exam I Study Guide
Note Weekly Reviews on
web, weeks 1-6. Note much of this info is ID to Study Guide, but some new things are there.
What are the main elements used by living
organisms? What are the rationales for life depending on H, O,
N, & C? Why add S & P? And how about the elements occurring
mainly as ions (Na, K, Ca, Cl, etc., & Fe, Zn, Cu etc.)? Why
should life be mainly a phenomena of the 2nd and 3rd electron
shells? (Think about this but don't spend too much time on it
on my account!) What are the main families of biomolecules and
what are their characteristic functional groups and functions?
Ionization; ion product; hydrophilicity - what
does this mean? Hydrophobicity - what does this mean? acid/base
properties of water. How can H+ move through water
apparently faster than the rate of diffusion? Structure of liquid
water. How does water structure affect solubility of other molecules?
hydrophobicity? Weak Bonds: What is a weak bond? What are the different types? (van der Waal's and Hydrogen bonds). Compare the relative strengths and stabilities of covalent, ionic, van der Waal's, and H-bonds. Be able to discuss these different bond types.
How are prokaryotes and eukaryotes distinguished? What are the various organelles we discussed and what happens in each? (basic biochemically important pathways, etc.)
Origin and Early Evolution of Life
How old is the Earth? How long ago did the Precambrian end? What are some of the major occurances important to life in the Precambrian? (origin, procaryotes, eucaryotes, multicellularity, etc.).
Know general acid/base properties, appearance
of titration curve, approximate values of pKa's and
ionic forms predominating at any pH. Know general formula for
aa's. Know which amino acid side chains are: hydrophobic, hydrophilic,
neutral, charged, polar.What's special about proline? Chirality
of aa's. Why is this significant? D & L. How many aa's are
used to synthesize proteins? Others are found in proteins - what's
happening? Memorize structures for: gly, ala, asp,
lys, cys, ser, leu, met, glu, phe.
Residue. Peptide bond (= amide bond); stability in
aqueous solution; planar nature of bond (resonance). Calpha, rotation angles - many are forbidden.
What is a globular protein?
Levels of description (Primary - Quaternary)
Periodicity, clustering, patterns. "Random" structure.
"Random" folding regions.
What are the characteristics of each level (residue relations,
bonding types, steric relations).
- Hydrophobic forces
- Hydrogen bonds
- Ion pairs = salt linkages
- Van der Waal's bonds
- Which are the most important bonds/forces at each level?
- Cooperativity effects in bonding.
- The effect of the aqueous environment on bond strength.
- Periodic peptide structures: alpha & beta (be able to describe these structures).
- What are some properties of alpha & beta structures?
- Why are they so common? (ease of nucleation, peptide H-bonds are satisfied). What aa(s) disrupt them?
- Remember there are two beta structures.
- Are these the only periodic structures found in proteins?
- The only extensive ones?
- The only extensive ones in globular proteins?
Supersecondary Structure/ Motifs
- What is the difference between a fold and a motif?
- What are some common motifs we've looked at/are in your text?
- What are some common folds we've looked at/are in your text?
Give examples to illustrate these concepts:
Hinges and domains.
Binding site/active sites are often split between domains.
Domain types (e.g. all alpha, alpha/beta, alpha+ beta, random)
What's the difference between a domain type and a motif?
Or is there one? Do the definitions overlap?
Domains, split active sites, genes and adaptation
- example of antibodies:
What is IgG?
How many chains does it have?
Does it represent a tertiary or quaternary structure? Defend
How many domains ?
How are the domains related?
How do we believe the domains arose?
Where are the active sites?
Assume your genome codes for 100 light chains and 100 heavy
chains. How many different IgG molecules can you make (without
mutations in the hypervariable region)?
IgG has been touted as a model for the evolution of advanced
proteins in eukaryotes. Explain.
Describes the overall folding of a single covalent structure.
Disulfide bonds - when are they formed, what are they good for, do they help in folding (as process - no), extra - vs. intracellular proteins.
Be able to discuss a protein's structure in terms of hierachical
levels and functional units/segments.
- What are the two structural "families" of fibrous
- 1 - made up of fibers much like a rope, e.g. collagen, keratin, silk fibroin.
- What are the special properties of keratin and silk fibroin?
- What's special about collagen:
- The collagen triple helix
- Sequence (periodicity of primary structure)
- Why so many gly?
- Why pro?
- 2 - made up of globules much like a string of 'snap beads'
(or analogous to a chain).
- e.g. microtubules, microfibrilles (actin fibers).
- Why quaternary structure?
- List and explain advantages.
- How does it occur?
- why do proteins often agregate/precipitate with heat?
- Chemical agents
- Be able to discuss urea as a denaturant.
- Why does it work (mechanisms)?
- When is a disulfide reducing reagent needed? Why only for
- Generally the lowest global DG
is not attained in folding proteins.
- What is argument for this statement?
- What is meant by a local DG minima?
- How do we envision a protein finding a stable structure?
- Folding pathways and nucleation.
- a-Helices, b-strands
- a-helix - b-strand
associations (super-2°/motif formation) and stabilization.
- Folding vs. rate of translation. Is there a temporal effect
in folding large proteins? Explain.
- Why domains in folding?
Chaperon Proteins (see also Discussion, below)
- What is a "heat shock" protein?
- What is a "chaperonin"?
- What is it used for?
- How do we think chaperonins work?
- Why are they needed?
Myoglobin/Hemoglobin and Binding
- Be able to discuss these proteins as oxygen storage/carrier
- Be able to discuss these proteins as examples of the topics
we have discussed under the general rubric of protein structure
- Describe the 1°, 2°, motif, domain, 3°, and 4°
structures. (Note, not all levels may be represented, or separately
- Note the heme group, how it is held, and by which subunits.
- Note the designation of the Hb tetratmer as an a2b2 or aabb
- If a protein had 8 subunits, with 3 of one kind, duplicates
of two kinds, and a singlet, how would you disignate this structure?
- Why is Hb called a dimer of dimers?
- What are the dimers?
- How are the monomers held in the dimers? (In general terms,
- What kinds of bonding predominate?
- How are the dimers held together?
- What kinds of bonding predominate?
- Be able to discuss these proteins as examples of binding
phenomena and its interpretation in proteins.
- When do we see a rectangular hyperbolic curve?
- What kind of a shape is this (be able to sketch it)?
- What kind of binding does it signify?
- What kind of binding (mathematical) model underlies it?
- What is a sigmoidal curve?
- What do we mean by cooperativity?
- What is the meaning of n in cooperative proteins?
Allosterism and Allosteric Enzymes
- What do V vs. [S] plots look like?
- Explain why this shape should arise from cooperative behavior (think about concentrations and multiple collision frequency/probability).
- What is meant by cooperativity?
- Describe a 100% cooperative system vs. a partially cooperative system
- What can we say about a system that exhibits a cooperativity of 2.5 ?
- Negative effector
- Negative effector vs. inhibitor (can be considered a special case).
- How does a (-) effector affect a V vs. [S] plot for a allosteric system? Why?
- What effect does does a (-) effector have on cooperativity? Why?
- Positive effector
- How does a (+) effector affect a V vs. [S] plot for a allosteric system? Why?
- What effect does does a (+) effector have on cooperativity? Why?
- Concerted (symmetry) model for allosteric enzymes.
- Be able to explain model (shifting equil.) for substrates (homotropic ) and effectors - correlate to kinetics.
- Be able to explain model (shifting equil.) for effectors (heterotropic) - correlate to kinetics.
- Sequential Model for allosteric enzymes. - explain.
What is an enzyme? (define) Turnover number. velocity.
Lock and Key model and its failure. Induced
fit model - explain. How do substrates bind? Chemical specificity.
Why are enzymes big (<5% of surface is active site)?
Zymogens: What are they? Why do they exist? (What enzymes commonly
occur as zymogens?)
What are main assumptions in steady-state derivation of the Michaelis-Menten
Note the consequences of the M-M eqn at:
- [S] >> Km,
- [S] << Km,
- [S] = Km.
- Be able to identify/explain reaction order at various [S].
Be able to interpret the Michaelis-Menten (vi vs. [S]) and Lineweaver-Burke (double-reciprocal) plots for both uninhibited
and inhibited reactions. Be able to find and/or show on/with both plots:
- Vmax and
- Km and apparent Km.
Know the three type of classical, reversible Inhibition
competitive: model mechanism, kinetics, plots;
non-competitive: model (mechanism), kinetics, plots;
un-competitive: model (mechanism).
Be able to draw and interpret plots of:
rate vs. temperature
rate vs. pH.
- What do vi vs. [S] plots look like?
- Concerted (symmetry) model for homotropic allosteric enzymes.
- Be able to explain model (shifting equil.) for substrates
and effectors - correlate to kinetics.
- Sequential Model - explain.
Know major types we discussed
- general acid/base (as opposed to specific),
- distortion/transition state binding,
- transition state charge stabilization,
- metal ion.
What is meant by a "concerted" mechanism?
Be able to explain an enzyme mechanism in terms of the catalytic
types we have discussed.
Be familiar with the mechanisms of lysozyme and the catalytic triad of the Serine proteases as we saw in class.
- Given the substrates and catalysts,
- be able to explain them in catalytic terms,
- be able to show reasonable electron movements for bond making/breaking
There will be one or two questions from discussion on the exam
- Be able to argue for or against the prion proposal.
- Why has the concept of Prions been so controversial?
- The prion diseases most closely resemble viruses. One way
to conceive of viral infections is as an introduction of foreign
(and disruptive) information into the host system (thus the use
of the term "virus" for invasive information in computer
- What "kind" of information does the author postulate
the prion transmits?
- How is this information transmitted/inherited?
- What evidence is there that only proteins are involved?
- What is thought to happen to the prion protein during transmission
of the disease?
- There seem to be genetically transmitted prion diseases (or
predilections to prion diseases). How do these diseases fit within
the Prusiner model?
Chaperons and Protein Folding
- What is the function of the chaperons? Is their existence
and necessity in many cases consistent with the idea that ALL
of the folding information for proteins is in the primary sequence?
- Why might protein aggregation be a major problem for cells
without chaperons? How do Chaperones prevent aggregation?
- What are the different types of chaperones (describe) and
how do their functions differ?
- Reconcile (or refute) the fact that chaperonins require ATP
energy for their activity with the statement that proteins "self-assemble."
- Is the suggestion of co-translational domain folding consistent
with a requirement for chaperons? Explain.
HIV and AIDS
- Consider the actions of the various drugs-what they are doing.
Be able to describe in terms of enzyme inhibition.
- Consider the viral responses (types of mutations, enzymes
involved) to the drugs..
- Consider the conclusions from this research, and the supplemental
(evolutionary) articles regarding:
- how HIV responds to challenges
- why HIV always "wins".
You may bring a data/information sheet to the exam,
however you must not exceed one side
of a single sheet of 5.5" x 8.5" paper (half
of a standard sheet of paper) for this sheet! GOOD LUCK!
Last modified 25 February 2011