UNIT 2 ENZYMOLOGY Flashcards
(190 cards)
1
Q
MAJOR ENZYMES
A
ACP
ALT
ALP
MS
AST
CK
GGT
LD
LPS
2
Q
MINOR ENZYMES
A
G6PD
PChE
OCT
ALS
LAP
3
Q
ATP: creatine-N-phosphotransferase
Physiologic function in muscle cells
82,000 daltons
Malaki yung size niya
associated with ATP regeneration
A
CK
4
Q
Fastest to migrate is the _____ , also known as CK-1
A
CK-BB
5
Q
CK-2 (hybrid type typically found in heart)
Suggestive of AMI
A
CKMB
6
Q
Normal level = below 6% of total CK
If more than 6%, it is suggestive of AMI
A
CKMB
7
Q
Slowest and most abundant CK
94-100% is
A
CKMM
8
Q
These are CK that are normally not present
A
atypical ck
9
Q
bound to the exterior surface of the inner mitochondrial membranes of muscle, brain, and liver migrate cathodal to CK-MM
0.8%-1.7%
A
mitochondrial ck
10
Q
indicator of severe illness, tx damage, and detected in cases of malignant tumor and cardiac abnormalities
A
mitochondrial ck
11
Q
migrate midway between CK-MM and CK-MB
comprised of CK-BB complexed with IgG, some IgA or complexes of lipoproteins with CK-MM
age and sex-related
Usually present in women over 50 years old
0.8%-1.6%
A
macro-ck
12
Q
Highest elevation of CK is caused by ______
A
Duchenne-type muscular dystrophy
13
Q
may visualize AK (cathodal to CK-MM)
A
electrophoresis
14
Q
Not an isoenzyme of CK but interferes with it. Causes falsely elevated results. Sumasama siya sa CK-MB.
A
AK= adenylate kinase
15
Q
have no CK but are rich in AK
A
RBC
16
Q
more sensitive and precise if done properly
Nagpapatong patong yung enzymes if done improperly
A
ion exchange chromatography
17
Q
coupled with PK-LD-NADH system; optimum pH of 9.0.
Forward method of CK
A
TANZER GILVARG
18
Q
coupled with HK-G-6-PD- NADP system; 2 to 6x faster; optimum pH of 6.8
Reverse method
A
OLIVER ROSALKI
19
Q
MW: 128,000 daltons
Class 1 eznyme
There’s oxidation-reduction reaction
Oxidoreductase enzymes participate in reactions wherein there is the transfer of hydrogen
Catalyzes the interconversion of lactic and pyruvic acids by transferring hydrogen using the coenzyme NAD+
A
LACTATE DEHYDROGENASE
20
Q
Tissue sources: heart, liver, skeletal muscle, kidney, and erythrocytes, lungs
A
LACTATE DEHYDROGENASE
21
Q
14-26%
A
LD-1
22
Q
29-39%
A
LD-2
23
Q
20-26%
A
LD-3
24
Q
8-16%
A
LD-4
25
6-16%
LD-5
26
normal healthy individual = dapat highest fraction is
LD-2
27
If mas mataas yung LD-1 kesa sa LD-2 may
flipped pattern
28
mixture of phenazine methosulfate and nitroblue tetrazolium reacts with NADH to produce a blue-purple color
WACKER METHODS
BLUE PURPLE COLOR
29
_______ reaction: Lactate to pyruvate in the presence of LD
FORWARD
30
pyruvate to lactate
REVERSE
31
Reverse method
We measure the disappearance of NADH
3x faster, smaller sample volumes, and shorter reaction times
susceptible to Substrate exhaustion and loss of linearity
WROBLEWSKI LA DUE METHOD
32
serves as cosubstrate and is consumed during the course of the reaction
NADH
33
is a major organ of LD, especially LD-1 and LD-2
RBC
34
LD is unstable
should be analyzed immediately within
48 hours stored at 25c
35
is the most labile isoenzyme, loss of activity occurs more quickly at 4°C than at 25°C
LD-5
36
involved in the transfer of an amino group between aspartate and a-keto acid
Transfer of an amino group from aspartate to a-keto glutarate to form glutamate
AST
37
uses pyridoxal phosphate (vit. B6) as a coenzyme
AST
38
tissue sources: cardiac tissue, liver, skeletal muscle; kidney, pancreas, erythrocytes
AST
39
DECREASED AST
UREMIA
40
Most abundant
cell cytoplasm AST
41
Only increase if there is significant cellular damage, which also damages the mitochondria
mitochondrial ast
42
43
a coupled enzymatic reaction
change in A at 340 nm
KARMEN METHOD
44
decrease in absorbance is measured
optimal pH is 7.3-7.8
KARMEN METHOD
45
ketoacids is reacted to 2,4-dinitrophenyl-hydrazine (DNPH) to form ketoacid hydrazones
AST
REITMAN -FRANKEL
46
product: intense brown color measured spectrophotometrically at 505 nm
REITMAN-FRANKEL
47
FALSE DECREASED OF AST
mercury
cyanide
fluoride
48
onset of CK-MB
4-8 hrs
49
onset of AST
8-12 hrs
50
onset of LD
12-24 hrs
51
peak of CK-MB
12-24 hrs
52
peak of AST
24 hrs
53
peak of LD
72 hrs
54
Duration of elevation of CK-MB
2-4 days
55
duration of elevation of AST
5 days
56
duration of elevation of LD
10 days
57
58
Catalyzes the transfer of an amino group from alanine to α-ketoglutarate
ALT
59
acts as coenzyme of ALT
pyridoxal phosphate
60
tissue sources: liver and cardiac tissue
ALT
61
bilirubin, erythromycin, iproniazid, morphine and endogenous pyruvate
Because endogenous pyruvate can be added to the reaction, leading to false elevation
false elevation in ALT
62
heavy metals like mercurial diuretics in ALT
false decreased
63
hydrolysis of various phosphomonoesters
alkaline phosphatase
64
optimal pH of ALP
9.0 to 10.0
65
requires Mg2+ and Mn2+ as activators (inorganic)
ALKALINE PHOSPHATASE
66
major tissue sources of ALP
INTESTINE
LIVER
BONE (osteoblasts)
67
MINOR TISSUE SOURCES OF ALP
spleen
placenta
kidney
68
major liver band and fast liver band
LIVER
69
Fast liver band also known as
A1 band
70
Major liver band is frequently elevated if Total ALP is _______ because of liver disease
increased
71
Normally ___________ in pregnancy, growth spurts, growing children, adults >50 years old.
elevated ALP
72
73
What LD is more suggestive of liver disease
LD5
74
What is LD6
Alcohol dehydrogenase
75
Arteriosclerotic cardiovascular failure
LD6
76
Macromolecular LD
● May sumama na immunoglobulin
● Migrates between LD-3 and LD-4
atypicl LD
77
highest LD concentration
megalobastic anemia
78
We can differentiate the presence of intestinal
isoenzyme depending on the blood group and
secretor status of the individual
B ANS O AND SECRETOR
79
80
Highly likely to have intestinal
ALP in the blood lalo na if after ingestion ng fatty meal
blood types B AND O
81
apparently bound by RBCs of
■ Blood type ____has a lower ALP since it is bound to
RBC.
A - RBC
82
Not a type of placental ALP. They are abnormal ALP that
are found if there are cancers or carcinoma involving the
placenta or reproductive system
carcinoplacental ALP
83
ectopic production of an enzyme by malignant
tissue; detected in various carcinomas like lung,
breast, ovarian, and colon (highest in ovarian and
gynecologic cancers)
REGAN
OVARIAN AND GYNECOLOGIC
84
85
Used for monitoring treatment because it
disappears upon successful therapy
REGAN
86
After the cancer cells are gone, there are no more
production of carcinoplacental ALP.
TRUE
87
2 types of carcinoplacental ALP
REGAN
NAGAO
88
Variant of the Regan
■ Detected in metastatic carcinoma of pleural
surfaces and in adenocarcinoma of the pancreas
and bile duct
NAGAO
PANCREAS AND BILE DUCT
89
Regan and Nagao frequently occurs in ____ of cancer
patients
3-15%
90
major liver band and fast liver or α1 liver
liver isoenzyme
91
92
Fast liver migrates ____ to the major band
anodal
93
heat stable to the least
PLACENTA
INTESTINE
LIVER
BONE
94
after heating, the remaining activity is <20%, source
of ↑ ALP
BONE
BABA SA 20
95
after heating, the remaining activity is >20%, source
of ↑ALP is the liver.
LIVER
TAAS NG 20
96
may be used to differentiate the source of
increase ALP whether it is from the liver or the bone.
HEAT STABILITY
97
most heat stable
and resist heat denaturation at 65 C for 30 minutes
PLACENTAL ALP
REGAN
NAGAO
98
Normal most stable:
PLACENTA ONLY
99
Abnormal most stable:
REGAN AND NAGAO
100
Placental ALP, Regan and Nagao are the most heat stable
and resist heat denaturation at
65c for 30 minutes
101
Phenylalanine inhibits
intestinal 75%
placental 80%
102
nagao is also inhibited by
L-LEUCINE
103
Urea (2M) inactivates
bone 90% more than liver 60%
104
HIGHEST TOTAL ALP
DISEASE
paget’s disease
105
more on obstructive than in
hepatocellular disorders (10x ULN due to cholestasis);
hepatitis and cirrhosis (<3x ULN)
hepatobiliary disorders
106
what liver enzyme?metastatic carcinoma of the
liver and other hepatobiliary diseases; indicator of
obstructive liver disease ALP and GGT
FAST LIVER ISOENZYME
107
ROTOR SYNDROME
NORMAL
108
Non-pathologic cause of increased ↑ALP:
PERIODS OF BONE GROWTH
ADULTS MORE THAN 50
PREGNANCY
109
inherited condition of hypophosphatasia due to
the absence of bone isoenzyme resulting to inadequate
bone calcification
DECREASED ALP IF MY HYPOPHOSPHATASIA
110
B-glycerophosphate or
Na-glycerophosphate
BODANSKY
111
p-nitrophenyl phosphate
BOWERS- McCOMB
112
BODANSKY
b-glycerophosphate
113
bowers mccomb
p- nitrophenyl phosphate
114
How does hemolysis affect CK, LD, and ALP
readings
Answer: CK - Adenylate kinase, LD - LD1 & LD 2, ALP is 6x more
in the RBC than in the serum
115
hydrolysis of various phosphomonoesters at an acidic pH
(5.0)
Just like your ALP, but performs in acidic pH
ACID PHOSPHATASE
116
tissue sources of ACP
prostate, bone, liver, spleen, kidney,
erythrocytes, and platelets
117
investigation of rape cases
ACP
118
inhibits prostatic
ACP (ACP is measured before and after inhibition with
substrate)
tartrate
119
inhibits non prostatic ACP
copper
120
falsely decreases ACP because the sample
becomes alkaline
CO2
121
Transfer gamma glutamyl from gamma-glutamyl peptides to
amino acids, H2O, and other small peptides
GGT
122
Involved in peptide and CHON synthesis, regulation of tissue
glutathione levels, transport of AA across CM
GGT
123
is an antioxidant, which is important in
protecting RBCs from oxidative damage.
GLUTATHIONE
124
Tissue sources of GGT:
tissues of the kidney, brain, prostate,
pancreas, liver
125
canaliculi of the hepatocytes, where
_____ is predominant
GGT
126
Class 2 enzyme
Also monitors chronic alcoholism and abstinence from
alcohol
GGT
127
markedly elevated in biliary tract obstruction
Patients taking enzyme inducing drugs like warfarin,
Phenobarbital, phenytoin (4x ULN)
hepatobiliary disorders
128
present in large extent in the smooth ER and
therefore subjected to
GGT; hepatic microsomal induction
129
If ALP is high and GGT is normal, the problem is in the
bone
BONE
ALP HIGH
GGT N
130
If GGT is high and ALP is normal, the problem is in the
hepatobiliary
ALP N
GGT HIGH
131
production of p-nitroanilide (yellow) or
5-amino-2-nitrobenzoate is monitored at 405 nm or 410 nm,
respectively
SZASZ METHOD- PHOTO,ETROC ENZYME REACTION
132
substrate used in szasz method photometric enzyme reaction
l-y-glutamyl-p-nitroanilide
133
MW: 50,000-55,000 daltons
○ smaller that’s why its observed in urine
Breakdown of starch and glycogen with products glucose,
maltose, dextrin
amylase
134
starch
amylose
amylopectin
135
long, unbranched chain of glucose
molecules, linked by α,1-4 glycosidic bonds
amylose
136
branched chain polysaccharide with
α,1-6 linkages at the branched points
amylopectin
BRANCHED
137
similar to amylopectin but more highly branched
glycogen
138
Requires Ca2+ and Cl- ions for activation
amylase
ALP MG AND MN
139
Major Tissues of amylase
acinar cells of pancreas, salivary glands
140
Importance: digestion of starches
AMYLASE
141
rise 2-12 hours after onset of attack and peak at 24 hours
but return to normal within 3-5 days
acute pancreatitis
142
Level of AMS is ______units/dL (2.55x ULN)
250-1000 somogyi
AMYLASE
143
Level of AMS is <500 Somogyi units/dL
intra-abdominal diseases
<500 somogyi units/dL
144
AMS combines with Ig to form a large complex that is
not filterable by the glomerulus
■ There’s no urine amylase because di maka exit sa
sobrang laki.
○ elevated AMS because of reduction in normal
clearance and urinary excretion is low.
macroamylasemia
145
predominates in urine
P-TYPE
146
predominates in serum
S TYPE
147
is markedly elevated in acute pancreatitis
P3
148
Increased in mumps or parotitis
■ migrates faster than the P-type
S TYPE
149
is inhibited by wheat _____
S TYPE
GERM LECTIN
150
measures the time required to reach the chromic point
fast disappearance means high AMS activity
iodometric method/amyloclastic
151
Nababawasan kulay =
mataas ang amylase activity
IODOMETRIC / AMYLOCLASTIC
TIME
152
starch substrate is hydrolyzed by AMS to its constituent
carbohydrate molecule that has reducing property like
sugars amount of reducing sugar α AMS activity (reported in
Somogyi units: number of mg of glucose in 30 minutes at
37°C at specific assay conditions)
saccharogenic method
153
amount of the reducing sugar formed using the reduction
method.
○Measure the appearance of the product.
saccharogenic
154
employs a starch substrate with a chromogenic dye that
forms an insoluble dye-substrate complex
the higher the color intensity of the soluble dye-substrate
complex, the higher the AMS activity
chromolytic
155
measures the change in absorbance of NAD+ at 340 nm at
pH=6.9
Substrate: maltotetrase or maltopentoase
Indicator reaction: conversion of NAD to NADH monitored at
340 nm
coupled enzyme
156
what substrate is used in coupled enzyme
maltotetrase or maltopentose
157
ph in coupled enzyme of amylase
pH= 6.9
340nm
158
Morphine and other opiates for pain relief may falsely
elevated AMS
159
in reduces the activity of AMS
LIPID
160
Oxalates and citrates may falsely
decrease
161
Hydrolyzes the ester linkages of fats (dietary TG) to produce
alcohols and fatty acids
Tissue sources: pancreas, stomach, intestine
LIPASE
162
Clinically specific isoenzyme of lipase
L2
163
is more specific in acute pancreatitis kasi
walang salivary isoenzyme.
L2
164
More specific for pancreatic disorder than AMS
lipase
165
LPS elevations persist for
5 days
166
albumin or ionized calcium
activators
167
heavy metals like quinine
inhibitors
168
fats in solution create a cloudy emulsion
Indicator reaction: decreased turbidity is monitored as LPS
hydrolyzes the turbid substrate fat emulsion
turbidimetric enzyme reaction
169
what is the substrate used in turbidimetric enzyme reaction
olive oil or triolein
170
measures liberated fatty acids released
by alkaline titration after 24 hours incubation using
phenolphthalein (pink) as indicator
titration: cherry-crandall method
171
same as Cherry Crandall but uses as indicator
Color blue
sigma tetiz
thymolphthalein
172
Oxidation of glucose-6-phosphate to 6-phosphogluconate
or the corresponding lactone
G6PD
173
functions to maintain NADPH in reduced form
G6PD
174
is required to regenerate
SULFHYDRYL-CONTAINING PROTEINS like glutathione
NADPH
175
protects Hb from oxidation. If hemoglobin
oxidizes, hemolysis occurs and may damaged the cell
membrane.
GLUTATHIONE NEEDS NADPH BY G6PD
176
G-6-PD deficiency: may result to
drug induced anemia
177
Increased level of G-6-PD
MI, MEGALOBLASTIC ANEMIA
178
No elevations seen in hepatic disorders (not elevated even if
all other enzymes are elevated)
G6PD
179
Red cell hemolysate sample:
enzyme deficiency
180
Serum sample:
enzyme ELEVATION
181
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