AA Flashcards
Enzymes defects that inhibits the body’s ability to metabolize certain amino acids
PKU Tyrosinemia Alkaptonuria MSUD Isovaleric Acidemia Homocystinuria Citrullinemia Arginosuccinic Aciduria Cystinuria
Phenylketonuria
Absence of _
_ of urine
phenylalanine hydroxylase (PAH) Musty odor
Laboratory Tests Phen
Guthrie test
Microfluorometric assay
Semi quantitative bacterial inhibition assay
Uses phenylalanine to facilitate bacterial growth (____ and___).
Guthrie test
B. subtilis and β-2-thienylalanine
Type II Tyrosinemia
↓ Tyrosine aminotransferase
Type III Tyrosinemia
↓ 4-hydroxyphenylpyruvate dioxygenase
Alkaptonuria
Lack of–
–
Lack of homogentisate oxidase
↑ homogentesic acid in urine
Type I Tyrosinemia
–
↓ fumarylacetoacetate hydrolase
Maple Syrup Urine Disease \_\_\_ \_\_ _ - \_\_\_odor of urine
↓ branched-chain α-ketoacid decarboxylase
↑ Isoleucine
↑ Leucine
↑ Valine
Burnt sugar
Map Syr test
Modified Guthrie test
Modified Guthrie test
uses branched chain ___ to facilitate bacterial growth (containing bbbbb and cvcvc).
α-ketoacid
B. subtilis
4-azaleucine
Isovaleric Acidemia
Deficiency of vvvvvvv
vvvvvv
vvvvv odor
Deficiency of isovaleryl-CoA dehydrogenase
↑ Isovaleric acid
Sweaty feet odor
Homocystinuria
Lack of vvvv
vvvv
vvvv
cystathionine β- synthetase
↑ Homocysteine
Methionine
Homocystinuria test
Uses vvv to facilitate bacterial growth (containing B. subtilis and vvvv).
Modified Guthrie test
Uses methionine to facilitate bacterial growth (containing B. subtilis and L-methionine sulfoximime).
Type I citrullinemia
Lack of vvvvv (ASAS)
Type I citrullinemia
Lack of arginonosuccinic acid synthetase (ASAS)
Mutation of the gene that encodes for protein citrin
Type II citrullinemia
Argininosuccinic aciduria
Lack of
argininosuccinic acid lyase (ASL)
Defect in amino acid transport system
Inadequate reabsorption of cystine in the kidneys
Cystinuria
Disintegration of protein to amino acids
degrades extracellular proteins
intra
Lysosomal pathway
Cytosolic pathway
Levels of protein structure
4
Primary
Secondary
Tertiary
Quarternary
Amino acids in a specific sequence
Regularly repeating structures stabilized by hydrogen bonds between the amino acids within the protein
Overall conformation (fold) of the protein molecule Due to interaction of side chains (e.g. ionic)
Interaction of more than 1 protein molecule or subunits
Primary
Secondary
Tertiary
Quarternary
Classification by Protein Functions 10
Enzymes Hormones Transport proteins Immunoglobulins (antibodies) Structural proteins Storage proteins Energy Source Osmotic force Homeostasis Acid-Base Balance
Contain peptide chains composed of only amino acids.
May be globular(_ _ ) or fibrous ()
Simple Proteins
May be globular (hormone, enzymes, transport) or fibrous (structural)
Conjugated Proteins
4
Metalloprotein
Lipoprotein
Mucoprotein and glycoprotein
Nucleoprotein
With higher carbohydrate -
10%-40% carbohydrate -
Nucleic acids attached-
Mucoproteins or proteoglycans - Mucin
Glycoproteins - Haptoglobin and α1-antitrypsin
Nucleoproteins - Chromatin
Plasma Proteins 2
Albumin
Globulin α1-Globulins α2- Globulins β-Globulins γ-Globulins
Indicator of nutrition
Binds thyroid hormones (_ _)
Binds retinol-binding protein
Prealbumin
T3, T4
Binds bilirubin, steroids, fatty acids
Major contributor to _ _
Albumin
oncotic pressure
α1-Globulins
α1- Antichymotrypsin Antitrypsin Lipoprotein Fetoprotein Acid glycoprotein Inter-α-trypsin inhibitor Gc-globulin
α1 - Antitrypsin
and
Acute phase reactant (α1 - Acid glycoprotein
)
Protease inhibitor
α1 - Fetoprotein
Principal fetal protein
↑ spina bifida, ↓ - Down syndrome
Transport lipids (HDL)
α1 – Lipoprotein
Inhibits serine proteinases
-
Gc-globulin
α1 - Antichymotrypsin
Inter-α-trypsin inh.
-
Transports Vit. D and binds actin
α2-Globulins
3
Haptoglobins - Acute Phase reactant, Binds Hgb.
Ceruloplasmin
α2 - Macroglobulin
Ceruloplasmin
Contains __
↓ - _ _
Contains copper
↓ - Wilson’s disease. Menkes synd
Inhibits protease
α2 - Macroglobulin
β-Globulins
8
Pre-β-lipoprotein β-Lipoprotein Trasferrin Hemopexin β2-Microglobulin C4, C3, C1q complement Fibrinogen C-reactive protein
Transports lipids (VLDL triglyceride) Transports lipids (LDL cholesterol)
Pre-β-lipoprotein
β-Lipoprotein
Transport Iron,↑-IDA,↓ Hemochromatosis
Trasferrin
Acute phase reactant, Binds heme
Hemopexin
Component of HLA molecules
β2-Microglobulin
Immune response (Opsonins)
C4, C3, C1q complement
Precursor of fibrin clot
Fibrinogen
Acute phase reactants
Promotes phagocytosis
C-reactive protein
γ-Gamma-Globulins Immunoglobulin G Immunoglobulin A M EE
Antibodies Antibodies in secretions Antibodies in early response Antibodies (reagen, allergy) Surface antibody
Other Proteins 7
Myoglobin
Troponin (cTn)
Fibronectin Fetal fibronectin (fFN)
Cross-Linked
C-Telopeptides
β-Trace Protein
Cystatin C
Amyloid
Cellular interaction
Placental adherence to the uterus
↑ - Preterm labor and delivery
Fibronectin Fetal fibronectin (fFN)
Cardiac marker for acute coronary syndrome
Troponin (cTn)
Oxygen carrier in muscles
Cardiac marker (AMI)
↑ 2-3 hrs of onset, peak at 8-12 hrs
Myoglobin
Proteolytic fragment of collagen I
Marker of bone resorption
Cross-Linked
C-Telopeptides
Syn: Prostaglandin D synthase
Marker for CSF leakage
β-Trace Protein
Cysteine proteinase inhibitor
Marker for kidney function (GFR)
Cystatin C
Fibrous protein aggregates formed from alteration of β pleated sheats
↑ Amyloidoses
Amyloid
Total Protein Abnormalities
TAG
N down up
Hepatic Damage
Cirrhosis β-γ bridging
Hepatitis ↑ γ-globulins
down down up
Acute -
Chronic -
Infections
Acute - α1 , α2 globulins
Chronic - ↑ α1, α2 γ, globulins
down down N
Inadequate diet
Nephrotic syndrome
↑α2,β-globulins;↓γ-globulins
Down N Down
Immunodeficiency syndrome
All UP
Dehydration
Up and Up
Multiple myeloma
Monoclonal &
Polyclonal gammopathies
Method of Analysis Total Protein
Kjeldahl
Refractometry
Biuret
Dye Binding
Method of Analysis Albumin and Globulin
Salt Precipitation
Dye Binding (Methyl orange, HABA, BCG, BCP)
Electrophoresis (Coomassie blue)
TP Reference method. Assume average ____ of 16%
Kjeldahl
nitrogen content
Measurement of refractive index due to solutes in serum
Refractometry
Formation of violet-colored chelate between VVV ions and VVVV
Biuret
Cu2+
peptide bonds
Protein binds to dye and causes a spectral shift in the absorbance maximum
Dye Binding
Kjeldahl 3
Kjeldahlization
Ammonia measurement-Nessler’s reaction (HgI2/KI)
-Berthelot reaction
conversion of nitrogen to ammonia
Nitrogen - - NH3
Kjeldahlization – H2SO4
Nitrogen H2SO4 NH3
Nessler’s reaction
Ammonia + Nessler’s rgt ___ — (NH2Hg2I)
Ammonia + Nessler’s rgt Gum ghatti Yellow solution (NH2Hg2I)
Berthelot reaction
Ammonia + alkaline hypochlorite ___ –
Na nitroprusside
Indophenol blue
Biuret Composition – breaks the peptide bonds – keeps copper in solution – stabilizes cupric ions
Cupric ions
Tartrate salt
Potassium iodide
Dye binding 5
Bromphenol blue Ponceau S Amido black 10B Lissamine green Coomassie brilliant blue
Globulins are precipitated
Albumin in supernatant is quantitated by biuret reaction
Salt Precipitation
Dye Binding
4
Methyl orange
HABA
BCG (Bromcresol green)
BCP (Bromcresol purple)
Nonspecific for Albumin
Methyl orange
HABA
Many Interferences
Many Interferences (salicylates, bilirubin)
Sensitive, Most commonly used dye
Bromcresol green
Specific, Sensitive and Precise
Bromcresol purple
Proteins separated based on electric charge densities
Electrophoresis
Electrophoresis (support media)
Cellulose acetate/agarose gel
Electrophoresis
After separation, protein fractions are immersed in acid solution then stained by dyes
(Coomassie blue)
scenned in densitometer
Uses higher voltage couple with a cooling system and more concentrated buffer
Electrophoresis (High Resolution)
It is the separation of molecules which takes place in silica capillaries.
Capillary Electrophoresis
Separates proteins on the basis of pI.
Uses constant power and __ or _ which contains a pH gradient.
pH gradient: ___
polyacrylamide or agarose gel
3.5 to 10
