Methods and techniques for the
isolation of proteins or nucleic acids
How can we isolate a specific protein for
First consider the SOURCE of your protein!
From The Strand
Magazine The Valley
of Fear (Frank Wiles)
What are the genetic differences between male- and female-determining sperm in
animals with heterogametic males?
ANS: The male-determining sperm carries a Y chromosome; the female-determining
sperm carries an X chromosome.
Stat 305, Fall 2013
This test is open book, open note, open resource. You may use the internet, but you may
not work together or get help from any other person. The test is due at the beginning of
class (9:25 AM) Tuesday, November 5, 2013.
3.1 On the basis of Mendels observations, predict the results from the following crosses
with peas: (a) a tall (dominant and homozygous) variety crossed with a dwarf variety; (b)
the progeny of (a) self-fertilized; (c) the progeny from (a) cross
In the human karyotype, the X chromosome is approximately the same size as
seven of the autosomes (the so-called C group of chromosomes). What procedure could
be used to distinguish the X chromosome from the other members of this group?
Beginning of Earth
o Hot, Reducing environment(no oxygen , little hydrolysis)
o electrical storms
o atmosphere filled with H2 CH4, NH3, HCN, H20 some CO, CO2
o inorganic materials ex iron, oxygen, silicon, magnesium, nickel, sul
o Six key functions:
1) Compartmentalization by a lipid bilayer and selective
permeation of metabolites and products.
2) Gene storage and replication.
3) Gene expression (DNA mRNA protein: transcription
Reactions useful for structure/function analysis
Cyanogen bromide cleavage
o Met-sulfur is a good nucleophile and reacts well with CNBr:
there are two protein c-c-n
o Iminolactone is easily hydrolyzed in the presence of water to cleave th
Transporting across membranes
Mechanisms of transporting
Case study - Bacteriorhodopsin and
Lipids, membranes and membrane associated
Why nonpolar things can be
important in biology too!
Types of lipids: struc
Structure and function
DNA, RNA and nucleotides.
Why all the complex abbreviations?
(e.g. why do I have to memorize so much stuff I
could look up in a book?)
Some basic terminology
macromolecule- usually refers to any big
Water and the properties of buffers
The chemistry of non-covalent
interactions with an emphasis on water
and its properties
Non-covalent interactions terminology and
Single-Bond Dissociation Energies at 25oC in kcal/mole
Shamoo Keck Hall Rm. 332
Lecture 1: Introduction to Biochemistry
Things start off pretty typically.
Time scale of evolution using a single day analogy
12:01 A.M. Earth forms and begins to cool (4.6 bya)
the building blocks of proteins
and chemistry of the 20
Introduction to protein primary
The synthesis of proteins from amino acids onethird of what has been called the central dogma
Chemistry of peptides, proteins and nucleic
Reactions useful for structure/function
Met-sulfur is a good
nucleophile and reacts well
Iminolactone is easily
hydrolyzed in the presence
of water to cleave t
Secondary, tertiary and quaternary structure
Keratin, collagen and silk
Levels of structural organization
tertiary structure of a protein is determined by its
Globular proteins: Protein folding and
How do proteins fold?
compact spherical shaped molecules
have a strongly hydrophobic core
much more common than fibrous proteins
can have structural or enzymatic properties
Hemoglobin and myoglobin
Applying protein structure to
How do myoglobin and hemoglobin
reversibly bind oxygen?
(tetramer with 4 hemes)
Hb has four polypeptides that strongly
resemble Mb and so Hb
Lec 1a Enzyme kinetics Olson
Fundamentals of Enzyme catalysis
Three basic functions of enzymes
(the essence of life):
1. Catalysis - speeding up reactions that are
ordinarily very slow (chemical and kinetic
mechanisms (VMAX(obs), KM(obs).
Chemistry of proteins and amino acids
How to characterize the protein you just
worked so hard to purify!
Success at purifying a novel protein is just the
first step in understanding it!
What is its amino acid composition/sequence?
How can we identify the