Enzymology - Lab Flashcards
1
Q
activities in the body that involves enzyme
A
- glycolysis
- creation of heme in hemoglobin
- coagulation cascade
- beta oxidation
- gluconeogenesis
2
Q
proteins within cells
A
enzymes
3
Q
why enzyme is used as catalyst?
A
because they make the chemical reaction faster
4
Q
does glycolysis still happen even without enzyme?
A
yes, but it takes 300,000 years for the action to be completed
5
Q
general properties of enzyme
A
active site: water free
allosteric site: cavity other than the active site
6
Q
a molecule or a substance where an enzyme acts
A
substrate
7
Q
factors of enzymology
A
- enzyme concentration
- substrate concentration
- cofactors
- inhibitors: competitive, non-competitive, uncompetitive
- isoenzymes
- temperature
- storage
8
Q
non-protein that must bond to a particular enzyme before a reaction occurs
A
cofactors
9
Q
electrolytes that are commonly used as cofactor in enzymatic reactions
A
- magnesium
- chloride
- calcium
10
Q
temp where enzymes are activated
A
25 - 37C
11
Q
temp where enzymes are being denatured
A
38-50C
12
Q
temp where enzyme will be inactivated
A
60-65C
13
Q
does not compete bcos the inhibitor will bind to other site
A
non-competitive
14
Q
binds w/ the enzyme substrate complex (no product/reaction will happen)
A
uncompetitive
15
Q
enzymes w/ similar enzymatic activities but differ their structure and origin
A
isoenzyme
16
Q
what is a fixed time
A
reacts at a designate time
17
Q
multiple measurements of absorbance changes are made during the reaction
A
kinetic assay
18
Q
enumerate the cardiac enzymes
A
- Creatine-Kinase ATP-Creatine-N-Phosphotransferase
- Lactate dehydrogenase L-Lactate: NAD Oxireductase
19
Q
CK is an enzyme important in the?
A
regeneration of adenosine triphosphate
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
21
Q
CK is dimeric because?
A
has 2 different monomers, the “M” and “B”
22
Q
transfers phosphate group between creatine phosphate and ADP
A
creatine kinase
23
Q
creatine kinase is also known as?
A
ATP-Creatine-N-Phosphotransferase
24
Q
CK is most commonly used in the diagnosis of?
A
- AMI
- muscular dystrophy
- CNS disorders
25
normal values for CK in males and female
male: **25-90 IU/mL (0.42-1.51 mmol/L)**
female: **10-70 IU/mL (0.17-0.18 mmol/L)**
26
sample used for testing CK
**serum**
anticoagulants inhibit CK
27
isoenzymes of creatine kinase "ATP-Creatine-N-Phosphotransferase" (from fastest)
- CK BB "brain type" CK1
- CK MB "hybrid type" "CK2"
- Macro CK
- CK MM "muscle type" "CK3"
- CK-Mi "mitochondrial CK"
28
isoenzyme abundant in cardiac and skeletal muscle (major activity is in the heart)
CK-MM "muscle type" "CK3"
29
major isoenzyme in sera of healthy people
CK-MM "muscle type" "CK3"
30
CK-BB is __ type "CK-3" which is normally found in?
neonatal sera rare in adult serum
31
CK-BB elevates in
brain injury and carcinomas
32
this isoenzyme is with significant amount in the heart and most specific for myocardial damage
CK-MB CK-2 "hybrid type"
33
CK-MB "hybrid type" CK-2
elevates:
peaks at:
normalize in:
elevates: **4-8 hrs**
peaks at: **12-24 hrs**
normalize in: **48-72 hrs**
34
this isoenzyme is falsely elevated in hemolysis
CK-MB CK-2 "hybrid type"
35
Macro-CK is common in ___ migrates middle of ___ and ___
Macro-CK is common in **older woman** migrates middle of ** middle of MM and MB** ___.
36
CK-Mi is also known as
mitochonrial CK
37
migrates cathodal to CK-MM
mictochondrial CK (CK-Mi)
38
mitochondrial CK migrates cathodal to CK-MM which indicates ____
several illness
39
enumerate the methods for CK
- Tanzer and Gilvarg Assay - forward
- Oliver Rosalki - reverse
- Electrophoresis
40
forward method of CK
ph:
absorbance:
Tanzer and Gilvarg Assay
ph: 9
absorbance: 340nm
41
reverse method of CK
ph:
absorbance:
| CK Method
Oliver Rosalki - reverse
ph: 6.8
absorbance: 340 nm
42
what is electrophoresis?
**separation of charged compounds**
(ex: serum protein, hgb, isoenzyme)
43
cathode
movement towards negative region
44
anode
movement towards positive region
45
Lactate dehydrogenase is also known as?
L-Lactate: NAD oxidoreductase
46
a transferase that hastens the interconversion of ____
**Lactate dehydrogenase**
a transferase that hastens the interconversion of lactic acid and pyruvic acid
47
LDH is found in?
various tissue sources
48
LDH is composed of ____ of two types: "M" (A) and "H" (B)
4 peptide chains
49
highest levels of LDH are detected in what diseases?
pernicious anemia and hemolytic disorders
50
disease associated with *elevations of LDH* are?
- hepatic disorders
- AMI
- pulmonary infarct
- acute lymphoblastic leukemia
51
forming glucose from non-carbohydrate sources
gluconeogenesis
52
isoenzymes
- LDH1
- LDH 2
- LDH 3
- LDH 4
- LDH 5
- LDH 6
- LDH1 (HHHH)
- LDH 2 (HHHM)
- LDH 3 (HHMM)
- LDH 4 (HMMM)
- LDH 5 (MMMM)
- LDH 6
53
major isoenzyme in healthy people
most abundant and heat stable
LDH 2 (HHHM)
54
an arteriosclerotic cardiovascular failure marker
LDH6
55
these LDH can be found ___
- LDH1 =
- LDH 3 =
- LDH 4 =
- LDH 5 =
- LDH1 = heart, rbc, kidneys
- LDH 3 = lungs, pancreas, spleen, lymphocytes
- LDH 4 = skeletal muscle, liver, intestine
- LDH 5 = liver, skeletal muscle, intestine
56
LDH can be used as a cardiac marker for these diseases, enumerate
- myocardial infarction
- hepatitis
- hemolysis
- lung and muscle disorders
57
LDH in myocardial infarction
elevates:
peaks:
remains:
elevates: **12 - 24 hrs** after onset
peaks: **48 - 72 hrs**
remains: **10 days**
58
for emergency diagnostic, what cardiac enzyme u will check?
Creatine kinase since it is the one who elevates faster
59
for delayed diagnosis, what enzyme should be check
lactate dehydrogenase
60
enumerate the LDH method
- forward or direct:
- reverse or indirect:
- forward or direct: Wacker method
- reverse or indirect: Wroblewski method
61
a direct LDH method
pH:
absorbance:
**Wacker Method**
pH: 8.8
absorbance: 340 nm
| LDH Method
62
Wroblewski Ladue
pH:
absorbance:
pH: 7.2
absorbance: 340 nm
3x faster than wacker
| indirect/reverse method of LDH
63
what are the other cardiac markers?
- myoglobin
- troponin I
- brain-type natriuretic peptide
64
transport and stores oxygen to intracellular respiratory enzymes of contractile cells
myoglobin
65
other cardiac marker with high sensitivity
myoglobin
66
myoglobin markers of AMI
- elevates ___ after onset
- peaks at _____ hours
- normalizes ___
- elevates **2-3** after onset
- peaks at **8-12 hrs** hours
- normalizes **18-30 hrs**
67
troponin I is found in ___ and with greater cardiac specificity
myocardium
68
troponin markers of AMI
- elevates ___ after onset
- peaks at _____ hours
- normalizes ___
- elevates **3-6 hrs** after onset
- peaks at **12-18** hours
- normalizes **6 days**
69
what is brain-type natriuretic peptide?
not and enzyme but heart failure marker
70
amylase is also known as?
Alpha 1-4 Glucan 4 Glucohydrolase
71
considered as the smallest enzyme
**Alpha 1-4 Glucan 4 Glucohydrolase**
| **amylase**
72
AMYLASE
activators:
substrate:
activators: **calcium and chloride**
substrate: **starch or glycogen**
73
acinar cells of the pancreas releases ___
amylopsin
74
salivary glands releases __
ptyalin
75
amylase clinical significance
- acute pancreatitis
- intestinal obstruction
- cholecystitis
- acute appendicitis
76
AMYLASE
elevates:
peaks:
normalizes:
elevates: **2-12** hrs after onset
peaks: **24** hrs
normalizes: **3-5** days
77
measures reducing sugar from starch breakdown?
| amylase method
Saccharogenic
78
- Measure ____ activity by following **decrease in substrate concentration**
- Follows breakdown of starch
Amyloclastic
79
Indicator of amyloclastic
Iodide
80
uses dye to check amylase activity
Chromogenic
81
Lipase is also known as
Triacylglycerol acylhydrolase
82
Lipase needs the presence of?
Bile salts and colipase
83
- 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
Lipase
84
Lipase clinical significance
- Acute Pancreatitis
- Perforated or duodenal ulcer
- Intestinal obstruction
- mesenteric vascular obstruction
85
Cherry Crandal
reference method for Lipase
86
Cherry crandal utilizes __ for hydrolysis
Indicator:
Use **olive oil** for hydrolysis
Indicator: *Phenolphthalein*
| substrate: olive oil, triolein (result: fatty acid PINK)
87
Most lipase activity found in serum are usually from the ___ and some are secreted by ___.
Most lipase activity found in serum are usually from the **pancreas** and some
are secreted by **gastric and intestinal mucosa**
