BIOC12H3 Lecture Notes - Lecture 3: Acid Hydrolysis, Carboxypeptidase B, Size-Exclusion Chromatography

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jadedolphin901

4 Dec 2012

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Bioc12fall2012 lecture week 3: protein primary structure and analysis (chapter 5) Increasing ionic strength at first increases the solubility (salting in),then decreases it (salting out) Then electrostatic interaction between protein molecules will make the proteins precipitate. Ion exchange chromatography: binding = low salt, washing = low salt, elution = high salt. 2 cation exchanger: negatively charged matrix so it can bind positively charged molecules cm (carboxymethyl) and. Mono s: anion exchanger: matrix is positively charged so it can bind negatively charged molecules deae (diethylaminoethyl) and mono q. ** the matrix is usually made up of inactive polymers. Size- exclusion chromatography: usage of gel filtration, purification of protein from contaminant proteins, determination of protein size by using mw standard and assuming the globular size of the protein, desalting usually last step of purification. Affinity chromatography: you have to choose 1:1 binding pairs, binding =, washing and elution steps are involved commonly used affinity tags are 6xhis (bindin to.

Related Questions

1. The level of protein structure that is stabilized primarilyby non-covalent interactions between the side chains within asingle polypeptide chain is refered to as its:

A) primary structure

B) secondary structure

C) tertiary structure

D) quaternary structure

E) none of the above

2. Collagen, a strong fibrous protein found in connectivetissue, bone, and the extracellular matrix, has a structure inwhich three left-handed helices wind around each other to form aright-handed triple helix. The primary structure of collagen has arepeat unit of -(Gly-X-Pro)- Which of the following bonds stabilizethe collagen triple helix? (mark all that apply)

A) Hydrogen bonds between hydroxyproline residues

B) Hydrogen bonds between N-H and C=O

C) Ionic bonds between amino acid side chains

D) Hydrogen bonds between charged amino acids

3. The non-covalent forces that stabilize the Beta-strandconformation of peptides include:

A) H-bonds between amide C=O and amide NH

B) Van der Waals packing between the peptide backbone atoms

C) Van der Waals packing due to side chain interdigitation

D) Only A and B above

E) None of the above

4. What is the approximate length difference of a 22-kDasingle-stranded ?-helical protein segment vs. a 22-kDasingle-stranded ?-strand protein segment? Assume a mean residuemass of 110 Da. Show work.

5. Circular dichroism measurements have shown that the peptidebackbone of poly-l-lysine (KKKKKKK) adopts a random coilconformation at pH 7.0, but becomes ?-helical as the pH is raisedabove 10. This observation is known as the

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Question 401 pts

Kinetic energy is the stored energy that can be used for motion.

True

False

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Question 411 pts

When an electron is transferred from one atom to another, and the two atoms are then electrically attracted to one another, a(n) ________________ bond is formed.

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Question 421 pts

Changing the number of ____________ of an atom would change the chemical properties of the atom.

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Question 431 pts

Forming molecules and breaking down molecules in biological organisms usually requires the use of _________ to help the reaction proceed faster.

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Question 441 pts

Organic molecules have a core composed of .

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Question 451 pts

The building blocks of carbohydrates are

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Question 461 pts

Water molecules crossing a membrane from high to low concentration is .

active transport

cell fate

facilitated diffusion

osmosis

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Question 471 pts

Choose the membrane molecule responsible for the passage of polar molecules and ions into and out of the cell.

phospholipids

transmembrane channel proteins

cell surface proteins

carbohydrate chains

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Question 481 pts

The simplest cells are .

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Question 491 pts

Which of the following membrane bound organelles are found inside bacterial cells:

nucleus

organelles are not found in prokaryotic cells

mitochondria

chloroplasts

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Question 501 pts

Which of the following is a specialized components of the cell, is associated with the rough endoplasmic reticulum, and is responsible for making proteins.

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Question 511 pts

Proteins are sorted, modified, and packaged by the _____________ and later transported to the outside of the cell.

ribosomes

golgi bodies (golgi apparatus)

nucleus

mitochondria

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Question 521 pts

Dehydration synthesis is a process of linking two smaller subunits together to form a polymer. Which of the following statements below is true of dehydration synthesis?

Oxygen is consumed.

A water molecule is removed from the molecules.

A water molecule is added to the molecules

Carbon dioxide is given off.

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Question 531 pts

Proteins are made up of _________ held together by peptide bonds.

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Question 541 pts

Which of the following gives an amino acid its properties?

NH2

R Group or functional group

COOH

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Question 551 pts

The final three-dimensional shape of a protein that includes the bonding of two or more polypeptide chains is call its _____ structure.

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Question 561 pts

How do DNA and RNA differ?

all statements are differences between DNA and RNA.

DNA is double stranded while RNA is single stranded.

They have different sugars.

Thymine is present in DNA but not in RNA.

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Question 571 pts

Which of the following is a component of a DNA nucleotide?

phosphate

a nitrogen containing base

All are components of a nucleotide.

5-carbon sugar

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Question 581 pts

Cell membranes are made up of several different types of molecules. Select the membrane molecule below that is made up of a polar region and two non-polar fatty acid tails.

transmembrane proteins

carbohydrate chains

phospholipids

cell surface proteins

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Question 591 pts

Two organelles which are believed to have once been free-living bacterial cells are ____________ and ______________.

chloroplasts and mitochondria

golgi apparatus and endoplasmic reticulum

ribosomes and nucleolus

peroxisomes and lysosomes

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