Chapter 13 - Enzymes Flashcards
these are rxn catalysts
enzymes
are all enzymes proteins?
yes
the greek word of enzyme means
in yeast
primary structure refers to
sequence of amino acids
secondary structure refers to
alpha helix
beta pleated sheets
tertiary structure is
folding/3D
kelan may quaternary ang protein?
> 1 polypeptid
substance acted upon
substrate
found in enzyme; where substrate reacts
active site
found in enzyme; for regulation, inhibition etc
allosteric site
same function with the enzyme but in different form
isoenzyme
how are isoenzymes differentiated
electrophersis
solubility
heat stability
chemical inhibition
isoenzymes are different in ______ structure and same in _______ structure
primary; tertiary
non-CHON necessary for enzymatic activity
cofactors
type of cofactors
coenzyme
activator
organic type of cofactors
coenzyme
inorganic type of cofactors
activators
examples of activators
Mg Ca Cl Cu Co Fe
examples of coenzymes
biotin NAD thiamine pyrophosphate lipoic acid flavin
coenzyme bound tightly to enzyme
prosthetic group
enzyme portionbound to prosthetic group
apoenzyme
prosthetic group + apoenzyme
holoenzyme
inactive form of an enzyme
proenzyme
classification of enzymes
oxidoreductase transferases hydrolases lyases isomerases ligases
what is the first digit in a pn enzyme’s code
classification of enzyme
what is the second digit in an enzyme’s code
subclass
responsible for the nomenclature of enzymes
IUB-EC
standard name of enzymes can be
substrate + “ase”
reaction + “ase”
enzyme responsible for oxidation reduction
oxidoreductase
enzyme responsible for the transfer of shit i don’t know what shit
transferase
enzyme responsible for hydrolysis
di ko alam eh
enzyme responsible for the removal of a group without hydrolysis
lyases
enzyme responsible for the interconversion of geometric npositional optical isomers
isomerase
enzyme responsible for the joining of two substrates
ligases
only example of a ligase
glutathione synthetase
two ways to allow product formation
giving more energy
using enzymes
energy required to raise all molecules in 1 mole of a compound at a certain temp to the transition state of the peak of energy
activation energy
types of specificity
absolute
group
bond
stereoisometric
optimum pH for most enzymes
7.0-8.0
optimum temp for most enzymes
37 oC
at what pH does trypsin work
1.0
high temp ________ enzymes
denatures
low temp ________ enzymes
inactivates
for every _____ increase in temp, rxn is doubled
10 oC
this shows the relationship betweenvelocity and substrate conc
Michaelis-Menten constant
the maximum point in the Michaelis-Menten curve is also the
saturation point
the point kung saan no matter gaano kadaming enzyme ang ilagay there is no more reaksyon na mangyayari
saturation point
jk di kosure kung ganyan ba un
types of inhibition
competitive
noncompetitive
uncompetitive
inhibitor binds direly to active site, reversible
competitive
inhibitor binds to a site other than thrpe active site
noncompetitive
when is noncompetitive inhibition irresversible
if damage is brought to the catalytic site
inhibitor binds to ES complex itself
uncompetitive
increased substrate in uncompetitive inhibition would _____ inhibition
worsen
the enzyme combines with only one substrate
absolute specificity
enzyme combines with all substrates contain a particular chemical group
group specific
enzymes combines to substrates with certain chemical bonds
bond specificity
enzymes that predominantly combine with only one optical isomer of a certain compound
stereoisometric specificity
the rxn rate is directly proportional to substrate conc.
first order kinetics
how is enzyme activity measured? thru:
increase in product formation
decrease in substrate formation(conc ata hindi form)
decrease in coenzyme formation
increase in conc of altered shitshit
amount of enzyme that will catalyze the rxn if 1 micromole of substrate per minute under specified condition enzyme activity
IU (kung anuman un)
two methods of enzyme activity measurement
fixed time/endpoint
continuous monitoring/kinetic assay
how many times is absorbance read in fixed time/endpoint
once
what are the major enzymes
TMTM
minor enzymes
OCT POD GOD ALS LAP
the recommended (?) name of creatine kinase
ATP:creatine-N-phosphotransferase
size of CK
82000 da
CK is associated mainly with _______ generation
ATP
major tissue sources of CK
skel muscle
heart
brain
minor tissue sources of CK
LLPPS PG BTUK
lung liver pancreas prostate spleen placenta GIT bladder thyroid uterus kidney
isoenzyme of CK
CK-MB
CK-MM
CK-BB
which CK isoenzyme is fastest toward anode
CK-BB
most electronegative CK isoenzyme
CK-BB
isoenzyme rarely seen in serum, mainly found in brain
CK-BB
least concentration among the CK isoenzymes
CK-BB
which CK isoenzyme is a hybrid
CK-MB
CK isoenzyme found mainly in the heart
CK-MB
CK isoenzyme found mainly in the skeletal muscle
CK-MM
normal CK-MB conc in total CK
<6%
two atypical isoenzyme of CK
mitochondrial CK
macro CK
where does mitochondrial CK migrate
before CK-MM (before origin)
CK isoenzyme indicating severe illness
mitochondrial CK
CK detected in cases of malignant tumor and cardiac abnormality
mitochondrial CK
where does ,acro CK migrate to
between MM and MB
this isoenzyme is CK-MM complexed with IgG, some IgA or lipoprotein
macro CK
age-sex related isoenzyme of CK
macro CK
normal conc. of macro CK
.8-1.6%
methods of CK isoenzyme measurement
electrophoresis
ion exchange chromatography
immunoassays
double antibody inhibition
what is visualized in electrophoresis of CK that is cathodal to CK-MM
AK (kung ano man yan)
immunoassay of CK isoenzymes use antibodies against ____________
H and B subunit
methods of CK det
Tanzer-Gilvarg
Oliver Rosalki
optimum pH of Tanzer Gilvarg
9.0
CK det method coupled w/ PK-LD-NADH system
Tanzer Gilvarg
forward type of CK det method
Tanzer Gilvarg
optimum pH of Oliver Rosalki
6.8
Ck det method coupled w/ HK-G6PD-NADP system
Oliver Rosalki
which is faster? Tanzer Gilvarg or Oliver Rosalki?
Oliver Rosalki
ref range of CK in males
15-160 U/L
CK-MB conc of >6% indicates
myocardial infarction
ref range of CK in females
15-130 U/L
instability of CK is resolved with
sulfhydryl compounds
example of sulfhydryl compounds
dithiothreitol
thioglycerol
does light inactivate CK
yes
higher CK values in men are attributed to incresed
muscle mass
enzyme responsible for interconversion of lactic and pyruvic acid
lactate dehydrogenase
how does LD work
transfer H+ using coenzyme NAD+
major tissue source of LD
heart
liver
skel muscle
erythrocytes
minor tissue sources of LD
lung
smooth muscle
diagnostic significance of LD
cardiac hepatic skel disease renal pernicious anemia
isoenzymes of LD
LD1 - LD6
fastest LD isoenzyme
LD1
highest conc among LD isoenzyme
LD2
highest affinity to anode LD isoenzyme
LD1
an LD idornzyme that is a bad prognosis and indicates impending death
LD6
order of LD isoenzymes most con to lowest conc
2 > 3 > 1 > 4 > 5
an atypical LD isoenzyme
macro LD
found between LD3 and LD4
macro LD
LD1 conc higher than LD2 indicates
myocardial infection
highest affinity LD isoenzyme to anode
LD1
methods for LD measurement
Wacker
Wroblewski La Due
endpoint of Wacker method
blue purple color
mol weight of LD
128000 da
LD6 is called
alcohol dehydrogenase
which LD measurement methd is forward
Wacker
by how many tomes is Wroblewski La Due faster than Wacker
3x
other name for AST
serum glutamic oxaloacetatic transaminase
major tissue sources of AST
cardiac muscle
skel muscle
liver
minor tissue sources of AST
kidney
pancreas
RBCs
methods for AST det
Reitman-Frankel
coupling with diazonium salts
mechanism of Reitman-Feankel
ketoacids is reacted to 2,4-dinitrophenylhydrazine (DNPH)
product of Reitman-Frankel
ketoacid hydrazones
falsely decreases AST level
mercury
cyanide
fluoride
example of liver diseases
viral hepatitis
cirrhosis
is AST useful in diagnosis of AMI
hindi eh
example of skeletal muscle disorder
muscular dystrophies
the optimum pH for Karmen method
7.3-7.8
absorbance of AST Karmen method
340nm
AST activity is stable in serum for ______ days at ref temp
3-4 days
catalyzes transfer of an amino group from alanine to alpha ketoglutarate wih the formation of glutamate and pyruvate
alanine aminotransferase
old term for ALT
serum glutamic pyruvic transaminase
major tissue source of ALT
liver
cardiac tissue
diagnostic significance of ALT
hepatic parenchymal disease
AMI or heart failure
IM
muscular dystrophy
most labile LD isoenzyme
LD5
LD activity in serum is _______
unstable
how is sample for LD stored
25 oC
gaano katagal dapat matest ang LD
within 48hrs
gaano katagal dapat matest ang LD isoenzyme
24hrs
ref range of LD
125-220 U/L
half life of ALT
24hrs
indicator enzyme for ALT assay
LD
absorbance for ALT
340nm
methods for ALT measurement
Karmen
Reitman
coupling w/ diazonium salt
unaffected by hemolysis
ALT
stability of ALT
3-4days at 4 oC
ref range for ALT
6-37 U/L
catalyzes hydrolysis of various phosphomonoesters at an alkaline pH
alkaline phosphatase
coenzyme for ALP
pyridoxal phosphate
activator for ALP
Mn
Mg
optimal pH for ALP
9.0-10.0
major tissue source of ALP
intestine liver bone spleen placenta kidney
ALP isoenzymes
liver ALP
bone ALP
placenta ALP
intestinal ALP
diagnostic significance of ALP
hepatobiliary
bone disorders
in bone disorders, there is elevation in ALP when ____ are involved
osteoblast
example of bone disorder where ALP is elevated
Paget’s disease
which ALP isoenzyme migrates the fastest
liver ALP
liver is further divided into
major liver band
fast or alpha 1
who are most likely to possess intestine ALP
blood type B and O
secretors
ALP isoenzyme differentiation
electrophoresis
heat stability
chemical inhibition
order of ALP isoenzymes in electrophoresis
fast > major > Regan Nagao > bone > placenta > intestinal
which ALP isoenzyme resust denaturation at 65 oC for 30 minutes
placental ALP
atypical ALP isoenzyme
Regan Nagao
other name for Regan Nagao
carcinoplacental ALP
ectopic product of an enzyme by malignant tissue
Regan isoenzyme
may be considered a variant of Regan isoenzyme
Nagao
what inhibits Nagao
L-leucine
which best inhibits intestinal and placentaL ALP
phenylalanine
detected in metastatic carcinoma and adenocarcinoma
Nagao
mechanism of Bowers McComb (ALP)
p-nitrophenylphosphate -> p-nitrophenol + phosphate ion
absorbance sa Bowers McComb
405nm
high fat meals _______ ALP
increase by 25%
ref range of ALP
30-90 U/L
optimal pH for ACP
5.0
tests for ALP
Bodansky King-Armstrong or Cutman Gutman Bessey-Lowry-Brock Hudson Babson Reed Bowers McComb Sinowara Jones Reinhart
basic difference in ALP and ACP
pH
major tissue source of ACP
prostate
minor source of ACP
bone liver spleen kidney erythrocytes platelets
diagnostic significance of ACP
hyperplasia of prostate
rape cases
bone disease
heart cancer
prostatic carcinoma
metastatic carcinoma
most specific substrate for prostatic ACP
thymolphthalein monophosphate
chemical inhibition of prostatic ACP usually uses ____ as inhibitor
tartrate
formula for pACP
total ACP - ACP after tartrate inhibition
PSA stands for
prostate-specific antibody
inhibitor for non-pACP
copper
serum activity _____ within _______ if the sample is left at rm tmp unpreserved
decrease; 1-2hrs
ref range for ACP
0-35 ng/mL
upon clotting, serum for ACP det must be
separated immediately
