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CC2 lab semi finals > Enzymology - Lab > Flashcards

Enzymology - Lab Flashcards

1
Q

activities in the body that involves enzyme

A
  • glycolysis
  • creation of heme in hemoglobin
  • coagulation cascade
  • beta oxidation
  • gluconeogenesis
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2
Q

proteins within cells

A

enzymes

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3
Q

why enzyme is used as catalyst?

A

because they make the chemical reaction faster

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4
Q

does glycolysis still happen even without enzyme?

A

yes, but it takes 300,000 years for the action to be completed

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5
Q

general properties of enzyme

A

active site: water free
allosteric site: cavity other than the active site

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6
Q

a molecule or a substance where an enzyme acts

A

substrate

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7
Q

factors of enzymology

A
  • enzyme concentration
  • substrate concentration
  • cofactors
  • inhibitors: competitive, non-competitive, uncompetitive
  • isoenzymes
  • temperature
  • storage
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8
Q

non-protein that must bond to a particular enzyme before a reaction occurs

A

cofactors

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9
Q

electrolytes that are commonly used as cofactor in enzymatic reactions

A
  • magnesium
  • chloride
  • calcium
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10
Q

temp where enzymes are activated

A

25 - 37C

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11
Q

temp where enzymes are being denatured

A

38-50C

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12
Q

temp where enzyme will be inactivated

A

60-65C

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13
Q

does not compete bcos the inhibitor will bind to other site

A

non-competitive

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14
Q

binds w/ the enzyme substrate complex (no product/reaction will happen)

A

uncompetitive

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15
Q

enzymes w/ similar enzymatic activities but differ their structure and origin

A

isoenzyme

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16
Q

what is a fixed time

A

reacts at a designate time

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17
Q

multiple measurements of absorbance changes are made during the reaction

A

kinetic assay

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18
Q

enumerate the cardiac enzymes

A
  • Creatine-Kinase ATP-Creatine-N-Phosphotransferase
  • Lactate dehydrogenase L-Lactate: NAD Oxireductase
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19
Q

CK is an enzyme important in the?

A

regeneration of adenosine triphosphate

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20
Q

why CK is most commonly used in the diagnosis of AMI, muscular dystrophy, and CNS disorders?

A

bcos they are found in skeletal muscles, heart muscles, and brain tissues

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21
Q

CK is dimeric because?

A

has 2 different monomers, the “M” and “B”

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22
Q

transfers phosphate group between creatine phosphate and ADP

A

creatine kinase

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23
Q

creatine kinase is also known as?

A

ATP-Creatine-N-Phosphotransferase

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24
Q

CK is most commonly used in the diagnosis of?

A
  • AMI
  • muscular dystrophy
  • CNS disorders
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25
Q

normal values for CK in males and female

A

male: 25-90 IU/mL (0.42-1.51 mmol/L)

female: 10-70 IU/mL (0.17-0.18 mmol/L)

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26
Q

sample used for testing CK

A

serum

anticoagulants inhibit CK

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27
Q

isoenzymes of creatine kinase “ATP-Creatine-N-Phosphotransferase” (from fastest)

A
  • CK BB “brain type” CK1
  • CK MB “hybrid type” “CK2”
  • Macro CK
  • CK MM “muscle type” “CK3”
  • CK-Mi “mitochondrial CK”
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28
Q

isoenzyme abundant in cardiac and skeletal muscle (major activity is in the heart)

A

CK-MM “muscle type” “CK3”

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29
Q

major isoenzyme in sera of healthy people

A

CK-MM “muscle type” “CK3”

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30
Q

CK-BB is __ type “CK-3” which is normally found in?

A

neonatal sera rare in adult serum

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31
Q

CK-BB elevates in

A

brain injury and carcinomas

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32
Q

this isoenzyme is with significant amount in the heart and most specific for myocardial damage

A

CK-MB CK-2 “hybrid type”

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33
Q

CK-MB “hybrid type” CK-2

elevates:
peaks at:
normalize in:

A

elevates: 4-8 hrs
peaks at: 12-24 hrs
normalize in: 48-72 hrs

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34
Q

this isoenzyme is falsely elevated in hemolysis

A

CK-MB CK-2 “hybrid type”

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35
Q

Macro-CK is common in ___ migrates middle of ___ and ___

A

Macro-CK is common in older woman migrates middle of ** middle of MM and MB** ___.

36
Q

CK-Mi is also known as

A

mitochonrial CK

37
Q

migrates cathodal to CK-MM

A

mictochondrial CK (CK-Mi)

38
Q

mitochondrial CK migrates cathodal to CK-MM which indicates ____

A

several illness

39
Q

enumerate the methods for CK

A
  • Tanzer and Gilvarg Assay - forward
  • Oliver Rosalki - reverse
  • Electrophoresis
40
Q

forward method of CK

ph:
absorbance:

A

Tanzer and Gilvarg Assay

ph: 9
absorbance: 340nm

41
Q

reverse method of CK

ph:
absorbance:

CK Method

A

Oliver Rosalki - reverse

ph: 6.8
absorbance: 340 nm

42
Q

what is electrophoresis?

A

separation of charged compounds
(ex: serum protein, hgb, isoenzyme)

43
Q

cathode

A

movement towards negative region

44
Q

anode

A

movement towards positive region

45
Q

Lactate dehydrogenase is also known as?

A

L-Lactate: NAD oxidoreductase

46
Q

a transferase that hastens the interconversion of ____

A

Lactate dehydrogenase

a transferase that hastens the interconversion of lactic acid and pyruvic acid

47
Q

LDH is found in?

A

various tissue sources

48
Q

LDH is composed of ____ of two types: “M” (A) and “H” (B)

A

4 peptide chains

49
Q

highest levels of LDH are detected in what diseases?

A

pernicious anemia and hemolytic disorders

50
Q

disease associated with elevations of LDH are?

A
  • hepatic disorders
  • AMI
  • pulmonary infarct
  • acute lymphoblastic leukemia
51
Q

forming glucose from non-carbohydrate sources

A

gluconeogenesis

52
Q

isoenzymes
- LDH1
- LDH 2
- LDH 3
- LDH 4
- LDH 5
- LDH 6

A
  • LDH1 (HHHH)
  • LDH 2 (HHHM)
  • LDH 3 (HHMM)
  • LDH 4 (HMMM)
  • LDH 5 (MMMM)
  • LDH 6
53
Q

major isoenzyme in healthy people
most abundant and heat stable

A

LDH 2 (HHHM)

54
Q

an arteriosclerotic cardiovascular failure marker

A

LDH6

55
Q

these LDH can be found ___

  • LDH1 =
  • LDH 3 =
  • LDH 4 =
  • LDH 5 =
A
  • LDH1 = heart, rbc, kidneys
  • LDH 3 = lungs, pancreas, spleen, lymphocytes
  • LDH 4 = skeletal muscle, liver, intestine
  • LDH 5 = liver, skeletal muscle, intestine
56
Q

LDH can be used as a cardiac marker for these diseases, enumerate

A
  • myocardial infarction
  • hepatitis
  • hemolysis
  • lung and muscle disorders
57
Q

LDH in myocardial infarction
elevates:
peaks:
remains:

A

elevates: 12 - 24 hrs after onset
peaks: 48 - 72 hrs
remains: 10 days

58
Q

for emergency diagnostic, what cardiac enzyme u will check?

A

Creatine kinase since it is the one who elevates faster

59
Q

for delayed diagnosis, what enzyme should be check

A

lactate dehydrogenase

60
Q

enumerate the LDH method
- forward or direct:
- reverse or indirect:

A
  • forward or direct: Wacker method
  • reverse or indirect: Wroblewski method
61
Q

a direct LDH method
pH:
absorbance:

A

Wacker Method
pH: 8.8
absorbance: 340 nm

LDH Method

62
Q

Wroblewski Ladue
pH:
absorbance:

A

pH: 7.2
absorbance: 340 nm

3x faster than wacker

indirect/reverse method of LDH

63
Q

what are the other cardiac markers?

A
  • myoglobin
  • troponin I
  • brain-type natriuretic peptide
64
Q

transport and stores oxygen to intracellular respiratory enzymes of contractile cells

A

myoglobin

65
Q

other cardiac marker with high sensitivity

A

myoglobin

66
Q

myoglobin markers of AMI

  • elevates ___ after onset
  • peaks at _____ hours
  • normalizes ___
A
  • elevates 2-3 after onset
  • peaks at 8-12 hrs hours
  • normalizes 18-30 hrs
67
Q

troponin I is found in ___ and with greater cardiac specificity

A

myocardium

68
Q

troponin markers of AMI

  • elevates ___ after onset
  • peaks at _____ hours
  • normalizes ___
A
  • elevates 3-6 hrs after onset
  • peaks at 12-18 hours
  • normalizes 6 days
69
Q

what is brain-type natriuretic peptide?

A

not and enzyme but heart failure marker

70
Q

amylase is also known as?

A

Alpha 1-4 Glucan 4 Glucohydrolase

71
Q

considered as the smallest enzyme

A

Alpha 1-4 Glucan 4 Glucohydrolase

amylase

72
Q

AMYLASE

activators:
substrate:

A

activators: calcium and chloride
substrate: starch or glycogen

73
Q

acinar cells of the pancreas releases ___

A

amylopsin

74
Q

salivary glands releases __

A

ptyalin

75
Q

amylase clinical significance

A
  • acute pancreatitis
  • intestinal obstruction
  • cholecystitis
  • acute appendicitis
76
Q

AMYLASE

elevates:
peaks:
normalizes:

A

elevates: 2-12 hrs after onset
peaks: 24 hrs
normalizes: 3-5 days

77
Q

measures reducing sugar from starch breakdown?

amylase method

A

Saccharogenic

78
Q
  • Measure ____ activity by following decrease in substrate concentration
  • Follows breakdown of starch
A

Amyloclastic

79
Q

Indicator of amyloclastic

A

Iodide

80
Q

uses dye to check amylase activity

A

Chromogenic

81
Q

Lipase is also known as

A

Triacylglycerol acylhydrolase

82
Q

Lipase needs the presence of?

A

Bile salts and colipase

83
Q
  • Hydrolyze glycerol esters of long-chain fatty acids
  • Most lipase activity found in serum are usually from the pancreas and some
    are secreted by gastric and intestinal mucosa
A

Lipase

84
Q

Lipase clinical significance

A
  • Acute Pancreatitis
  • Perforated or duodenal ulcer
  • Intestinal obstruction
  • mesenteric vascular obstruction
85
Q

Cherry Crandal

A

reference method for Lipase

86
Q

Cherry crandal utilizes __ for hydrolysis
Indicator:

A

Use olive oil for hydrolysis
Indicator: Phenolphthalein

substrate: olive oil, triolein (result: fatty acid PINK)

87
Q

Most lipase activity found in serum are usually from the ___ and some are secreted by ___.

A

Most lipase activity found in serum are usually from the pancreas and some
are secreted by gastric and intestinal mucosa

CC2 lab semi finals (7 decks)

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