Enzymes: Lactate Dehydrogenase and Creatine Kinase Flashcards

1
Q

What reaction does Lactate Dehydrogenase catalyze?

A

The oxidation of lactate to pyruvate or pyruvate to lactate

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

How many pH optimums does the Lactate Dehydrogenase reaction have?

A

2

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

What is the pH optimum for the forward LD reaction?

A

8.3-8.9

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

What is the pH optimum for the reverse LD reaction?

A

7.1-7.4

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

What is the required co-factor for the LD reaction?

A

NAD+

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

The LD reaction can occur in both directions, which direction does this reaction favor?

A

Reverse (pyruvate to lactate)

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

Is lactate dehydrogenase specific for lactate and pyruvate?

A

Not absolute specificity. Can also reduce a number of alpha ketoacids and oxidize several alpha hydroxyacids

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

What is the molecular structure of lactate dehydrogenase?

A

Tetramer (4 polypeptide chains) bound together by disulphide bonds

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

What are the 2 possible LD subunit types?

A

H and M subtypes

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

What are some LD inhibitors?

A

Borate, oxalate, EDTA, pyruvate, lactate, reagents with reactivity against thiol groups

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

How do borate and oxalate inhibit LD enzymatic activity?

A

They compete with lactate for the enzyme binding site

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

How does EDTA inhibit LD enzymatic activity?

A

It binds with zinc, which acts as an LD activator.

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

How do pyruvate and lactate inhibit LD enzymatic activity?

A

They inhibit due to excess

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

Why would serum LD be increased?

A

Heart disease, liver disease, muscle disease, hemolytic disorders, pulmonary disease, cancer

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

Name H and/or M subunits of LD-1.

A

H4

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

Name H and/or M subunits of LD-2.

A

H3M

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

Name H and/or M subunits of LD-3.

A

H2M2

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

Name H and/or M subunits of LD-4.

A

HM3

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

Name H and/or M subunits of LD-5.

A

M4

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

How many LD isoenzymes are there?

A

5 isoenzymes

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

Which LD isoenzyme(s) will be increased when heart disease is present?

A

LD-1 and LD-2

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

Which LD isoenzyme(s) will be increased when liver disease is present?

A

LD-4 and LD-5

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

Which LD isoenzyme(s) will be increased when muscle disease is present?

A

LD-5

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

Which LD isoenzyme(s) will be increased when hemolytic disorders are present?

A

LD-1 and LD-2

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
25
Which LD isoenzyme(s) will be increased when pulmonary disease is present?
LD-3
26
Which LD isoenzyme(s) will be increased when cancer is present?
LD-4 and LD-5
27
What type of sample is preferred for LD measurement?
Serum
28
Why isn't plasma preferred when LD measurement is done?
May be contaminated with platelets, which contain a high concentration of LD
29
Does hemolysis affect LD measurement?
Yes, it will falsely elevate the levels due to LD being present in RBCs
30
Which LD isoenzymes are labile to cold?
LD-4 and LD-5
31
What is recommended storage for serum used for LD measurement?
Keep at room temp because some of the LD isoenzymes are sensitive to cold
32
In children, are LD levels normally higher or lower than adults?
5x higher
33
Measurement of LD depends on what?
Whether the reaction is occurring in a forward or reverse direction
34
Why is the LD forward reaction the recommended way to measure it?
Lactate is more stable than pyruvate as a substrate, less inhibitory
35
What is actually measured when LD enzymatic activity is assayed?
As lactate is oxidized to pyruvate, NAD is reduced to NADH. There will be an increase of absorbance at 340 nm.
36
Why would LD isoenzymes be fractionated?
To determine the organ of involvement in the disease process
37
Which isoenzyme can be isolated using substrate selectivity with 2-oxybutyrate?
LD-1, due to it containing only H subunits, which 2-oxybutyrate has a greater affinity for
38
Immunoprecipitation is used to measure all LD isoenzyme fractions except for?
LD-1
39
Which enzyme group does creatine kinase belong to?
It is a transferase
40
What reaction does CK catalyze?
Catalyzes the reversible transfer of a phosphate group from ATP to creatine
41
What is pH optimum for forward CK activity?
9.0
42
What is the pH optimum for reverse CK activity?
6.8
43
What ion does CK require to activate the reaction?
Mg
44
What compounds can act as inhibitors on CK?
(Excess) Mg, Ca, Zn, Cu, sulfhydryl binding reagents, citrate, fluoride, uric acid, excess ADP
45
What is the molecular structure of CK?
It is a dimer; 2 peptide chain, B and M joined by disulphide bonds
46
How many CK isoenzymes are there?
3 (CK-1, CK-2, CK-3/CK-BB, CK-MB, CK-MM)
47
How is CK relatively unstable?
Activity is easily lost when the thiol group in the disulphide bond is broken.
48
Can the disulphide bond in the CK molecule be restored?
Partial restoration can occur by adding sulfhydryl compounds
49
What tissue sources is CK-1 (CK-BB) mainly contained in?
Brain and nerve tissue
50
What tissue sources is CK-2 (CK-MB) mainly contained in?
Heart and skeletal muscle
51
What tissue sources is CK-3 (CK-MM) mainly contained in?
Heart and skeletal muscle
52
What are CK isoforms?
Subtypes of the CK isoenzymes
53
How are CK isoforms created?
Arise from the removal of the terminal lysine on the M peptide chain of the isoenzymes released from the tissues
54
What enzyme catalyzes the removal of the terminal lysine from the M peptide chain on the CK molecule?
Carboxypeptidase
55
How many isoforms does CK-3 (CK-MM) have?
3
56
Which CK-3 isoform has both lysines intact?
MM3
57
Why doesn't carboxypeptidase have any effect on the CK-1 (CK-BB) isoenzyme?
The enzyme only has an effect on the M peptide chain. The CK-1 isoenzyme has only B peptide chain.
58
Which CK-2 (CK-MB) isoform does not have lysine removed?
MB2
59
Which CK-2 (CK-MB) isoform has one lysine removed?
MB1
60
How long does it take to remove the lysine from the M peptide chains to form the CK isoforms?
Takes about 10 hours
61
Which CK-3 isoform has only one of the lysines removed?
MM2
62
Which CK-3 isoform has both lysines removed?
MM1