Brainscape's Knowledge GenomeTM

Browse over 1 million classes created by top students, professors, publishers, and experts.


See full index

Biochemistry > Protein Purification and Characterization Techniques > Flashcards

Protein Purification and Characterization Techniques Flashcards

1
Q

What tends to happen to the percent recovery during a protein’s purification?
a. The number usually steadily increases during the purification.
b. The number usually steadily decreases during the purification.
c. The number usually stays fairly constant during the purification.
d. There is no general trend for percent recovery during a protein purification.

A

b

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

Differential centrifugation at low speeds (several thousand RPM) is a useful step when
a. organelles have been lysed.
b. enzymes of interest have different sizes.
c. cell membranes must be left intact.
d. ribosomes need to be broken down.
e. there are either organelles or debris to separate.

A

e

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

The typical order for the major steps of enzyme isolation would be (from first to last):
a. Homogenization, salt fractionation, electrophoresis, column chromatography.
b. Homogenization, column chromatography, salt fractionation, electrophoresis.
c. Homogenization, salt fractionation, column chromatography, electrophoresis.
d. Salt fractionation, homogenization, electrophoresis, column chromatography.
e. Homogenization, electrophoresis, salt fractionation, column chromatography.

A

c

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

The purity of an enzyme at various stages of purification is best measured by
a. total protein.
b. total enzyme activity.
c. specific activity of the enzyme.
d. percent recovery of the protein.
e. percent recovery of the enzyme.

A

c

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

Salting out with ammonium sulfate is based upon proteins interacting with other proteins via
a. hydrogen bonds.
b. ionic bonds.
c. hydrophobic interactions.
d. disulfide bonds.

A

c

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

Using differential centrifugation it is possible to separate
a. nuclei, mitochondria, and ribosomes into three separate fractions
b. organelles from contaminating salts
c. proteins that differ in charge
d. proteins from membranes

A

a

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

The following methods are useful for cell homogenization:
a. Sonication.
b. Freezing and thawing.
c. Detergents.
d. Enzymes.
e. All of these are correct.

A

e

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

Which of the following happens as a protein is purified?
a. the percent recovery and the fold purification both increase
b. the percent recovery and the fold purification both decrease
c. the percent recovery increases and the fold purification decreases
d. the percent recovery decreases and the fold purification increases

A

d

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

Ammonium sulfate is useful in protein purification because
a. it contains nitrogen and sulfur, both of which occur in proteins
b. it is sparingly soluble in water, causing proteins to co-precipitate with it
c. very pure proteins are obtained when it is used
d. it forms ion-dipole interactions with water, making proteins less soluble and more likely to precipitate

A

d

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

The typical order of differential centrifugation for organelles is (from slowest speed/lowest g to fastest speed/highest g):
a. nuclei, microsomes, mitochondria & chloroplasts, cytosol, whole cells
b. whole cells, nuclei, mitochondria & chloroplasts, microsomes, cytosol
c. cytosol, microsomes, nuclei, mitochondria & chloroplasts, whole cells
d. nuclei, mitochondria & chloroplasts, whole cells, cytosol, microsomes
e. whole cells, cytosol, microsomes, nuclei, mitochondria & chloroplasts

A

b

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

Which of the following are principles on which to base column chromatography?
a. Molecular size
b. Isoionic pH or pI
c. Ion exchange
d. Both molecular size and ion exchange
e. All of these

A

e

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

Which separates on the basis of molecular weight?
a. Gel filtration
b. Affinity chromatography
c. Cation exchange
d. Anion exchange
e. Cation or anion exchange

A

a

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

Which separates based on the ionic charge on a protein?
a. Gel filtration
b. Affinity chromatography
c. Cation exchange
d. Anion exchange
e. Cation or anion exchange

A

e

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

Which type of column is most affected by the shape of the protein, for example, comparing spherical and cigar-shaped
proteins?
a. Gel filtration
b. Affinity chromatography
c. Cation exchange
d. Anion exchange
e. Cation or anion exchange

A

a

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

Which would be best to separate a protein that binds strongly to its substrate?
a. Gel filtration
b. Affinity chromatography
c. Cation exchange
d. Anion exchange
e. Cation or anion exchange

A

b

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

Elution of proteins by means of a pH gradient would work best with this type of column:
a. Gel filtration
b. Affinity chromatography
c. Cation exchange
d. Anion exchange
e. Cation or anion exchange

A

e

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
17
Q

Which would be best to separate positively charged proteins?
a. Gel filtration
b. Affinity chromatography
c. Cation exchange
d. Anion exchange
e. Cation or anion exchange

A

c

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
18
Q

Which would be best to separate proteins of similar size?
a. Gel filtration
b. Affinity chromatography
c. Cation exchange
d. Anion exchange
e. Cation or anion exchange

A

e

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
19
Q

In affinity chromatography, a protein
a. which binds to the ligand will remain on the column.
b. which binds to the ligand will elute from the column.
c. which is hydrophobic will remain on the column.
d. which is hydrophilic will remain on the column.

A

a

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
20
Q

In gel filtration chromatography
a. materials are separated based on their size, the smaller ones eluting first.
b. materials are separated based on their size, the larger ones eluting first.
c. materials are separated based on their hydrophobic nature, the more hydrophobic ones eluting first.
d. materials are separated based on their hydrophobic nature, the less hydrophobic ones eluting first.

A

b

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
21
Q

The order of elution of AAs H, E, & K from a cation exchange column by a pH 6 buffer is
a. H E K
b. E H K
c. K H E
d. E K H

A

b

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
22
Q

In any form of chromatography, how will a compound which interacts more strongly with the stationary phase elute compared to one that interacts less strongly?
a. A compound interacting more strongly will elute earlier than one with weaker interactions.
b. A compound interacting more strongly will elute later than one with weaker interactions.
c. The order of elution has nothing to do with interactions with the stationary phase, but with interactions with
the mobile phase.

A

b

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
23
Q

A chromatography technique where a solution of nonpolar compounds is put through a column that has a nonpolar
liquid immobilized on an inert matrix is which type of chromatography?
a. Gel filtration
b. Ion exchange
c. Affinity
d. HPLC
e. Reverse Phase HPLC

A

e

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
24
Q

In a sample consisting of lysine, leucine, and glutamic acid, which will be eluted last from an anion exchange resin at
pH 7?
a. all three will be eluted at the same time
b. lysine
c. leucine
d. glutamic acid

A

d

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
25
Q

In chromatography the experimental setup always requires
a. a stationary phase and a mobile phase
b. a spectrophotometric detecting device
c. a sample in which components differ in charge
d. a sample in which components differ in polarity

A

a

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
26
Q

A separation of a mixture of cations of different charge requires
a. another cationic substance
b. an anionic substance
c. an electrically neutral, but highly polar, substance
d. an electrically neutral, nonpolar substance

A

b

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
27
Q

Which of the following is not an example of column chromatography?
a. ammonium sulfate fractionation
b. ion-exchange separation
c. HPLC
d. affinity separation

A

a

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
28
Q

In affinity chromatography
a. there is nonspecific binding of proteins to column material
b. only minor purifications can be obtained
c. the mobile phase is always pure water
d. the ligand is always specific for one type of protein to be bound
e. there can be molecule specific ligands or group specific ligands

A

e

29
Q

The degree of separation in molecular sieve chromatography depends on
a. the polarity of the mobile phase
b. the pKa of the buffer material in the mobile phase
c. the chemical nature of the sieve material
d. the size of the pores in the sieve material

A

d

30
Q

In all forms of chromatography one way of identifying eluted substances is by
a. fluorescence spectroscopy
b. comparison with standards
c. radioactive labeling
d. treating fractions with a reagent that will cause a color change

A

b

31
Q

Which substance would you expect to be eluted first from a molecular sieve column with a suitable degree of crosslinking?
a. hemoglobin
b. myoglobin
c. 2,3-bisphosphoglycerate
d. all would elute at the same rate

A

a

32
Q

In the SDS-PAGE (sodium dodecylsulfate - polyacrylamide gel electrophoresis) method, separation takes place on the
basis of
a. charge only, because all particles have different charges, but the same mass.
b. the sieving action of the gel, because all particles have the same charge, but different masses.
c. the sieving action of the gel, because all particles have approximately the same charge/mass ratio, but different
masses.
d. the chemical nature of the buffer used as the electrolyte.

A

c

33
Q

How many bands would be produced when hemoglobin is subjected to SDS-PAGE?
a. 1
b. 2
c. 3
d. 4
e. 2, but only if the size of the pores in the gel would allow two proteins of slightly different size to be separated

A

e

34
Q

The following are all principles on which to base electrophoresis except:
a. Molecular size
b. Isoionic pH or pI
c. Net charge
d. Binding to a substrate
e. Shape

A

d

35
Q

Two-dimensional electrophoresis usually exploits these 2 different properties of proteins:
a. Molecular weight and shape
b. Molecular weight and net charge
c. Molecular weight and isoionic pH
d. Isoionic pH and shape
e. Isoionic pH and net charge

A

b

36
Q

When electrophoretic separations are done based on molecular weight, the distance that a molecule moves can be
graphed as a straight line when compared to:
a. the MW of the proteins
b. the negative of the MW of the proteins
c. the log of the MW of the proteins
d. none of these

A

c

37
Q

In electrophoresis experiments
a. the separation must be carried out in bright light
b. the polarity of substances to be separated is more important than their charge or size
c. the sample can be badly degraded as a result of the separation
d. an electric field must be applied to the mixture to be separated

A

d

37
Q

In isoelectric focusing gel electrophoresis
a. particular care must be taken to ensure the same pH along the length of the gel
b. there is a pH gradient that parallels the electric field gradient
c. the electric current is allowed to fluctuate
d. the electric circuits of the apparatus must be very well insulated

A

b

38
Q

The isoelectric point is
a. the pH at which a substance has no net charge
b. the pH at which a substance has a net positive charge
c. the pH at which a substance has a net negative charge
d. the pH at which a substance has no charge groups of any kind

A

a

39
Q

Two dimensional separation methods
a. lead to unreliable results
b. are not widely used because of their complexity
c. do not improve separation
d. consist of two separation methods applied in sequence

A

d

40
Q

Important steps in sequencing pure proteins include all of these except:
a. Determining the amino acid composition
b. Determining the isoionic pH of the protein
c. Breaking the protein into smaller peptides
d. Determining the amino acids on the ends of the protein
e. Determining the amino acids on the ends of the smaller peptides

A

b

41
Q

If a protein with the sequence FEWPRQVDMARINE is treated with chymotrypsin, what will the products be?
a. F EW PRQVMARINE
b. FE WPRQVD MARINE
c. FEWPR QVDMAR INE
d. FEWPRQVDM ARINE

A

a

42
Q

Which of the following treatments results in a polypeptide fragment with a homoserine lactone at the C-terminal end?
a. trypsin
b. chymotrypsin
c. cyanogen bromide
d. Edman method

A

c

43
Q

If a protein with the sequence PQRKYPIG is treated with trypsin, what will the products be?
a. PQR KYPIG
b. PQRK YPIG
c. PQR K YPIG
d. PQ R KPIG0

A

c

44
Q

Cyanogen bromide (CNBr) cleaves proteins
a. after positively charged residues, such as K & R.
b. after negatively charged residues, such as D & E.
c. after aromatic residues, such as Y & W.
d. after methionine residues.

A

d

45
Q

It is impossible to sequence a protein if you do not have overlapping sequences to work with.
a. True
b. False

A

b

46
Q

The most efficient method for determining the sequence of a short peptide is:
a. Edman degradation
b. Trypsin digestion
c. Chymotrypsin digestion
d. Cyanogen bromide digestion

A

a

47
Q

Methods for breaking proteins into smaller peptides include all of the following except:
a. Digestion with chymotrypsin
b. Cyanogen Bromide treatment
c. Digestion with Trypsin
d. Edmann degradation
e. All of the above create short peptides suitable for sequencing

A

d

48
Q

When end-group analysis was done on the protein insulin, the results indicated that both glycine and phenylalanine
were N-terminal amino acids and both asparagine and alanine were C-terminal amino acids. These results indicate that
a. the experiment was done incorrectly
b. no conclusions can be drawn
c. there were impurities in the sample
d. insulin consists of two polypeptide chains

A

d

49
Q

Matrix-Assisted Laser Desorption Ionization is a type of _____ technique.
a. electrophoresis
b. ion exchange chromatography
c. affinity chromatography
d. mass spectrometry

A

d

50
Q

An amino acid analyzer is an instrument used to determine
a. the sequence of amino acids in a polypeptide chain
b. the identity of N-terminal and C-terminal amino acids in a protein
c. the presence of modified amino acids in a protein
d. the identities and relative amounts of amino acids in a protein

A

d

51
Q

Generally speaking, sequence techniques have become so sensitive that if you are able to isolate the protein on a gel,
there is enough of it to get a significant amount of its sequence.
a. True
b. False

A

a

52
Q

Determination of the sequence of amino acids in a peptide is done by
a. x-ray crystallography
b. Edman degradation
c. treatment with cyanogen bromide
d. trypsin hydrolysis

A

b

53
Q

It is frequently possible to bypass the determination of the identity of the N-terminal amino acid of a protein because
a. this information is already available from the amino acid analysis
b. the Edman method sequences the peptide from the N-terminal end
c. N-terminal amino acids are always chemically modified
d. this information is not needed

A

b

54
Q

Which of the following is not used in protein structure determination?
a. digestion with proteolytic enzymes
b. the Edman method
c. treatment with cyanogen bromide
d. treatment with alkyl halides

A

d

55
Q

In an ELISA, the molecule that binds directly to the target protein is called the
a. primary antibody
b. secondary antibody
c. linked enzyme
d. immunosorbent

A

a

56
Q

Which of the following explains why a secondary antibody is used in an ELISA?
a. two antibodies will be twice as visible as one antibody
b. it allows the visible signal to be amplified
c. primary antibodies cannot be tagged with visible markers
d. none of the choices

A

b

57
Q

In an ELISA, the compound 4-chloro-1-naphthol is used because
a. it helps the secondary antibody to bind to the protein
b. it helps the primary antibody bind to the protein
c. it turns color in the presence of an enzyme that is bound to the secondary antibody
d. all of the choices

A

c

58
Q

Which of the following is NOT true concerning a western blot?
a. it is usually performed in a 96 well microtiter plate
b. it involves the use of primary antibodies
c. it involves the use of secondary antibodies
d. it involves electrophoresis

A

a

59
Q

The purpose of using nitrocellulose for a western blot is to
a. allow the various proteins to migrate and separate
b. bind to the primary antibody
c. allow the proteins to be concentrated onto the face of the nitrocellulose where they will be accessible to the
primary antibody
d. to allow the 4-chloro-1-naphthol to react and be visible

A

c

60
Q

A protein chip is functionally most similar to which other technique?
a. gel filtration
b. ELISA
c. SDS-PAGE
d. western blot

A

b

61
Q

The most common way of visualizing the results seen on a protein chip is by use of
a. secondary antibodies
b. primary antibodies
c. fluorescence
d. using an enzyme-linked reaction involving 4-chloro-1-naphthol

A

c

62
Q

Studying the entire complement of proteins being produced by a cell is called
a. genomics
b. proteomics
c. ELISA
d. western blot

A

b

63
Q

Studying how proteins interact with other proteins is called
a. expression proteomics
b. structural proteomics
c. interaction proteomics
d. none of the choices

A

c

64
Q

The process of breaking cells open to release the organelles is called _____.
a. comminution
b. distillation
c. homogenization
d. liquefaction

A

c

65
Q

Which of the following polysaccharides is used to make up resins for use in electrophoresis and in column
chromatography?
a. Fructan
b. Agarose
c. Callose
d. Glucan

A

b

66
Q

What is the distinguishing feature of affinity chromatography?

A

The distinguishing feature of affinity chromatography is that the polymer is covalently linked to
some compound, called a ligand, that binds specifically to the desired protein. The other proteins in the sample do not bind to the column and can easily be eluted with buffer, while the bound protein remains on the column.

67
Q

Which of the following is a similarity between ion-exchange chromatography and affinity chromatography?
a. Both use a column resin that binds the protein of interest.
b. Both involve a less-specific interaction and are based on net charge.
c. Both separate molecules of varying molecular weights.
d. Both use very high-resolution columns that can run under high pressures

A

a

68
Q

Identify a proteolytic enzyme that preferentially hydrolyzes amide bonds adjacent to aromatic amino acid residues.
a. Lipase
b. Chymotrypsin
c. Amylase
d. Tyrosinase

A

b

Biochemistry (13 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2024 Bold Learning Solutions. Terms and Conditions