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Clinical Chemistry (CC 1) > (M) Lec 5: Amino Acids (P2: Proteins) > Flashcards

(M) Lec 5: Amino Acids (P2: Proteins) Flashcards

1
Q

Most plasma proteins are synthesized in what organ?

A

Liver

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2
Q

The liver is responsible for producing numerous plasma proteins except for what?

A

Immunoglobulins (aka antibodies which are produced by lymphocytes)

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3
Q

This panel is used to test the synthetic function of the liver (if it normally produces the proteins needed by the body)

A

Protein Panel

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4
Q

The process of degrading proteins to become free amino acids

A

Protein Catabolism

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5
Q

Protein Catabolism Pathways

Degrades extracellular and some intracellular proteins

A

Lysosomal pathway

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6
Q

Protein Catabolism Pathways

Important in degrading intracellular proteins

A

Cytosolic pathway

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7
Q
  • Refers to the removal of amino acid nitrogen from the body
  • Removal of an α-amino group (N alone is toxic to the body) from an amino acid by transaminases to a ketocarbon which produces ketoacids
A

Transamination

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8
Q

These 2 products of transamination:
1. Are converted to urea by the liver (via the urea cycle)
2. Are converted to glucose/fat (via the Krebs cycle)

(2 answers)

A
  1. Ammonia
  2. Ketoacids
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9
Q

Protein Structures

  1. Overall shape and confirmation; there is folding and a 3D configuration (determines the function and phsyiochemical properties)
  2. Alpha-helix and Beta-pleated sheet (adds strength and flexibility)
  3. The basic amino acid sequence
  4. The shape/structure from the interaction of one or more protein molecules/sub-units held by noncovalent bonds

A. Primary
B. Secondary
C. Tertiary
D. Quaternary

A
  1. C
  2. B
  3. A
  4. D
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10
Q

Protein Properties

TOF: Proteins are always negatively charged

A

False (can be BOTH)

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11
Q

Protein Properties

Refers to the pH at which amino acids and protein have no net charge

A

Isoelectric Point

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12
Q

Protein Properties

  • If pH > pI, a protein has a net (+ or -) charge
  • If pH < pI, a protein has a net (+ or -) charge
A
  • pH > pI = negative
  • pH < pI = positive
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13
Q

Protein Properties

Basic or Acidic Group?
Lysine, Arginine, and Histidine

A

Basic

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14
Q

Protein Properties

Basic or Acidic Group?
Glutamate, Aspartate, Cysteine, and Tyrosine

A

Acidic

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15
Q

Protein Properties

If there is a charge at the protein surface, the protein is (hydrophilic or hydrophobic)?

A

Hydrophilic

Note: The charge makes it more soluble

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16
Q

Protein Properties

A protein’s lowest solubility is at what point?

A

Its isoelectric point (there is no net charge)

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17
Q

Protein Classifications (by function)

  • These catalyze chemical reactions
  • Most are composed of proteins EXCEPT ribozyme (composed of nucleic acids)
A

Enzymes

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18
Q

Protein Classifications (by function)

Chemical signals that are important for the endocrine system

A

Hormones

Can be made up of proteins or steroids

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19
Q

Protein Classifications (by function)

These are responsible for carrying certain ions/molecules

A

Transport Proteins

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20
Q

Protein Classifications (by function)

  • Are antibodies important for the defense mechanism and immunity
  • Binds to foreign antigens
  • Not produced by the liver but rather the plasma cells
A

Immunoglobulins

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21
Q

Protein Classifications (by function)

Examples are fibers, collagen, and keratin

A

Structural Proteins

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22
Q

Protein Classifications (by function)

An example is ferritin (iron-storage)

A

Storage Proteins

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23
Q

Protein Classifications (by function)

A source of this comes from gluconeogenesis

A

Energy Source

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24
Q

Protein Classifications (by function)

  • Pressure that holds the fluid inside the blood vessels
  • Is greatly affected by albumin therefore this functions to distribute water throughout the different compartments of the body
A

Osmotic Force

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25
Q

Protein Classifications (by structure)

Composed purely of proteins and amino acids
- Globular (enzymes, messengers, transporters, hemoglobin, plasma proteins, and peptide hormones)
- Fibrous/Structural (fibrinogen, troponin, and collagen)

A

Simple

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26
Q

Protein Classifications (by structure)

Attached to non-protein molecules
- An apoprotein + prosthetic group
- Metalloproteins - ferritin, hemoglobin, and ceruloplasmin (carries copper)
- Lipoproteins - HDL, LDL, VLDL, and chylomicrons
- Glycoproteins - anti-trypsin and haptoglobin
- Nucleoproteins - chromatin
- Mucoproteins/Proteoglycans - mucin

A

Conjugated

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27
Q

Nitrogen Balance

  • Protein catabolism exceeds anabolism
  • Burns, wasting disease, high fever, and starvation
A

Negative Nitrogen Balance

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28
Q

Nitrogen Balance

  • Protein anabolism is greater than catabolism
  • Growth, pregnancy, and repair processes
A

Positive Nitrogen Balance

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29
Q

Plasma Proteins

What are the 2 major groups?

A

Albumin and Globulin

Majority is albumin (50-60%)

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30
Q

Plasma Proteins

Plasma Protein Panel Normal Values:
1. Total Protein
2. Albumin
3. Globulin
4. A/G Ratio

A. 1.1-1.8
B. 6.5-8.3 g/dL
C. A computed value only
D. 3.5-5.5 g/dL

A
  1. B
  2. D
  3. C
  4. A

Globulin = TP - Albumin

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31
Q

Plasma Proteins

What is the SI unit conversion factor for proteins?

A

g/dL x 10 = g/L

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32
Q

Serum Protein Electrophoresis

  • pH = ?
  • Charge of proteins = (+ or -)
  • Protein separation and migration move from what point of origin to what end point?

(3 answers)

A
  1. pH = 8.6
  2. Proteins are (-) charged
  3. From cathode (-) to anode (+)
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33
Q

Serum Protein Electrophoresis

Order the proteins from most to least abundant and their migratory distance:

  1. Y-globulins
  2. Albumin
  3. B-globulins
  4. A1-globulins
  5. A2-globulins

Note: Their concentrations and migratory distances are ordered the same

A
  1. Albumin
  2. A1-globulin
  3. A2-globulin
  4. B-globulin
  5. Y-globulin

Study the percentages in the recorded lecture

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34
Q

High Resolution Electrophoresis

Separated into as many as how many bands?

A

12

Note: Study the additional zones in the lecture

Additional modifications include: Higher voltage, cooling system, and a more concentrated buffer

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35
Q

Serum Proteins

  • Aka transthyretin; it migrates ahead of albumin
  • Transport protein for thyroxine (T4) and triiodothyronine (T3)
  • Binds with retinol binding protein to form a complex that transports retinol
  • Half life:** 2 days**
  • Rich in tryptophan
  • A sensitive marker of poor nutritional status
A

Pre-Albumin

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36
Q

Serum Proteins

  • Highest protein concentration in the plasma (50-65%)
  • Synthesized by the liver at 9-12 g/day
  • Half life: 20 days
  • Glycated Albumin - Fructosamine (alternative to HbA1c)
  • Functions:
    1. Maintains 80% of colloid osmotic pressure (a decrease leads to edema)
    2. Negative acute phase reactant (decreases when there is infection/inflammation)
    3. Buffers pH
    4. Transportation (Iron, Fatty Acids, Bilirubin (B1), Electrolytes, Steroids, Drugs, Hormones)
A

Albumin

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37
Q

Albumin

Increased (hyperalbuminemia) or Decreased (hypoalbuminemia)?
1. Inadequate source of amino acids in malnutrition and malabsorption
2. Liver disease
3. Protein losing enteropathy or gastrointestinal loss (diarrhea)
4. Kidney loss to urine in renal disease
5. Skin loss in burns or exfoliative dermatitis
6. Hypothyroidism
7. Dilution by excess intake of water or administration of intravenous fluids
8. Genetic mutation resulting from an autosomal recessive trait
9. Redistribution by hemodilution (e.g. sepsis)
10. Increased catabolism - massive burns and malignancy

A

Decreased (hypoalbuminemia)

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38
Q

Albumin

TOF: Hyperalbuminemia has a higher clinical significance than hypoalbuminemia

A

False (hyperalbuminemia has no significance)

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39
Q

Albumin

Increased (hyperalbuminemia) or Decreased (hypoalbuminemia)?
- Severe dehydration and prolonged torniquet application

A

Increased (hyperalbuminemia)

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40
Q

Albumin

  • An inherited error of albumin synthesis
  • Albumin level is decreased because the patient is not capable of synthesizing it
A

Analbuminemia

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41
Q

Albumin

  • Can be inherited or acquired
  • 2 albumin bands are present in electrophoresis
A

Bisalbuminemia

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42
Q

Serum Proteins

  • Consists of α1, α2, β and γ fractions (through electrophoresis)
  • Each consists of a number of different proteins with
    different functions
A

Globulins

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43
Q

Globulins

What are the 2 negative acute phase reactants?

A

Albumin and Transferrin

44
Q

Globulins

  • The most abundant alpha-1 globulin (90% of the fraction)
  • It neutrallizes trypsin-like enzymes: Elastase - a neutrophil protease (major inhibitor of protease activity)
  • Increases in inflammatory reactions, pregnancy, and contraceptive use
A

α1-Antitrypsin (AAT)

45
Q

Globulins

This enzyme degrades elastin, fights infections, and can also destroy alveoli which leads to emphysema

46
Q

Globulins

A mutation wherein the α1-Antitrypsin (AAT) is deficient/abnormal which causes:
- Juvenile hepatic cirrhosis
- Lung disease (emphysema); ZZ phenotype (the normal is MM)

Elastase is increased/not inhibited

47
Q

Globulins

  • Synthesized in the developing embryo and fetus and by parenchymal cells of the liver
  • Protects the fetus from** immunologic attack by the mother**
  • Is increased during pregnancy but will decrease gradually after birth, reaches adult levels by 8-12 months
  • No known function in normal adults
  • Tumor Marker (L3) for liver and testicular cancer
  • Screening between 15 to 20 weeks of gestation for Neural Tube Defects and Down Syndrome
  • Screening specimen: Maternal serum
  • **Confirmatory specimen: **Amniotic fluid
  • Interpretation requires accurate dating of pregnancy
  • Affected by maternal weight, race, and DM
A

α1-Fetoprotein (AFP)

48
Q

α1-Fetoprotein (AFP)

Elevated or Decreased AFP?
Spina bifida, neural tube defects, abdominal wall defects, anencephaly, HDN, and general fetal distress

49
Q

α1-Fetoprotein (AFP)

Elevated or Decreased AFP?
Increased risk for Down syndrome and Trisomy 18

50
Q

α1-Fetoprotein (AFP)

What are the 2 testing methods?

A

Radioimmunodiffusion (RID) and Enzyme-labeled Immunoassay (or ELISA)

51
Q

α1-Fetoprotein (AFP)

Migratory region in electrphoresis? (it lies between 2 other proteins)

A

Between Albumin and A1-Globulin

52
Q

Globulins

  • Aka Orosomucoid
  • An acute phase reactant
  • High percentage of CHO (45%) and sialic acid (11-12%)
  • Is negatively charged with **low pH **
  • Greatest affinity for progesterone and binds to Quinidine (cardioactive drug)
  • Diagnostic tool for neonates with bacterial infection
A

α1-Acid Glycoprotein

53
Q

α1-Acid Glycoprotein

Elevated or Decreased?
Pregnancy, stress, inflammation and tissue damage, acute
myocardial infarction, trauma, pregnancy, cancer, pneumonia, and rheumatoid arthritis and surgery

54
Q

α1-Acid Glycoprotein

What is the normal value during testing (RIA, immunofixation, and nephelometry)?

A

50-120 mg/dL

55
Q

Globulins

  • Serine proteinase inhibitor (serpin)
  • Inhibits the enzymes: cathepsin G, pancreatic elastase, mast cell chymase and chymotrypsin
  • Produced by the** liver**
  • An acute phase reactant (decreased in liver disease)
  • Mutations in patients with Parkinson’s disease and COPD
  • Increased in Alzheimer’s disease
A

α1- Antichymotrypsin

56
Q

Globulins

  • A serine protease inhibitor
  • Plays a role in inflammation and carcinogenesis
  • It migrates in the Alpha 1 and Alpha 2 interzone (hence the name); in conventional electrophoresis, it is still part of the A1 fraction
A

Inter-α-trypsin Inhibitor (ITIs)

57
Q

Globulins

  • aka Group-specific component globulin
  • Part of the Alpha 1 and Alpha 2 interzone
  • Carries vitamin D
  • Transports fatty acids and endotoxins
  • Binds with actin for clearing
  • Co-chemotactic factor for neutrophils and monocytes in inflammation
  • A prognostic indicator of survival with tissue injury after trauma and hepatic failure
A

Gc-globulin

58
Q

Gc-globulin

Elevated or Decreased?
During the third trimester of pregnancy and patients taking estrogen oral contraceptives

59
Q

Gc-globulin

Elevated or Decreased?
Liver disease and protein losing syndrome

60
Q

Gc-globulin

Decreased levels after trauma/hepatic failure indicate what prognosis?

A

Poor prognosis (low survival rate)

61
Q

Gc-globulin

What is the method of choice?

A

Immunonephelometry

62
Q

Globulins

  • An α2–glycoprotein synthesized in the hepatocytes
  • An acute phase reactant
  • Binds hemoglobin to prevent urinary loss
  • **3 phenotypes: **Hp1-1, Hp2-1 and Hp2-2 (increased risk for CVD in patients with DM)
  • For evaluation of** anemia** (hemolytic vs. other causes); evaluate the degree of intravascular hemolysis that occurs in transfusion reactions or HDN
A

Haptoglobin

63
Q

Haptoglobin

Is it indicative of hemolytic anemia?
Decreased haptoglobin with decreased reticulocyte count

A

No (reticulocytes should be increased)

64
Q

Haptoglobin

Is it indicative of hemolytic anemia?
Normal haptoglobin with increased reticulocyte count

A

Yes (RBC destruction in the spleen and liver)

65
Q

Haptoglobin

Is it indicative of hemolytic anemia?
Normal haptoglobin with normal reticulocyte count (BUT with signs of anemia)

A

No (a different cause of anemia other than hemolytic)

66
Q

Haptoglobin

Is it indicative of hemolytic anemia?
Decreased haptoglobin without signs of anemia

A

No (a problem with the synthetic function of the liver)

67
Q
  • An α2–glycoprotein for the transport of copper
  • Synthesized in the liver
  • An acute phase reactant
  • 90% or more of total serum copper is found in this, 10% bound in albumin
A

Ceruloplasmin

68
Q

Ceruloplasmin

  • An autosomal recessive condition wherein ceruloplasmin is low while tissue copper storage and urinary secretions are high (decreased levels in the blood)
  • Characterized by liver cirrhosis, CNS damage, and a Kayser-Fleischer ring in the cornea
A

Wilson’s Disease

Additional: Menkes Kinky Hair Syndrome is also a copper metabolism disorder

69
Q

Ceruloplasmin

Methods of Determination:
Early analytic method

A

Copper oxidase activity

70
Q

Ceruloplasmin

Methods of Determination:
Immunochemical method

A

Radial immunodiffusion and nephelometry

71
Q

Globulins

  • From the liver, a large protein (major component of α2 band)
  • Inhibits proteases such as trypsin, pepsin, thrombin, kallikrein and plasmin by trapping (a role in hemostasis)
  • Removal of reticuloendothelial cells
  • Forms a complex with PSA (prostate specific antigen)
A

α2-Macroglobulin

72
Q

α2-Macroglobulin

Increased or Decreased?
DM, Liver disease, Nephrosis, Pregnancy, and Contraceptive use

A

Increased

In nephrosis, it is 10x higher

73
Q

α2-Macroglobulin

Familiarize with the methods of determination

A

Immunodiffusion, immunonephelometry, ELISA, and latex agglutination immunoassay

74
Q

Globulins

  • A β-glycoprotein synthesized by the liver
  • A negative acute phase reactant
  • Most important for the iron pool; transports iron to prevent urinary loss and tissue deposition
  • Transports to storage sites (incorporated to apoferritin → ferritin)
  • Reversibly binds 2 molecules of ferric iron
  • Iron bound to transferrin is less than 0.1% (4 mg) of total body iron
A

Transferrin (Siderophilin)

75
Q

Transferrin (Siderophilin)

Increased or Decreased?
Impaired hemoglobin production leading to secondary iron-deficiency anemia

A

Decreased (iron is not recycled)

76
Q

Transferrin (Siderophilin)

Increased or Decreased?
Hereditary disorder in iron metabolism and hemochromatosis (iron in liver and pancreas)
- iron overload leads to **increased iron bound to transferrin **with tissue depletion (low transferrin)

77
Q

Transferrin (Siderophilin)

A hereditary condition wherein no transferrin leads to anemia and hemosiderosis

A

Atransferrinemia

78
Q

Transferrin (Siderophilin)

High or Low?
Iron Deficiency Anemia

79
Q

Globulins

  • β globulin
  • Synthesized by the liver
  • An acute phase reactant
  • A scavenger of heme (1:1 ratio) to protect the body from oxidative damage and it also preserves iron
  • Heme-Hemopexin complex is carried to the liver for destruction
80
Q

Hemopexin

High or Low?
Diagnostic marker of hemolytic anemia

81
Q

Hemopexin

High or Low?
Inflammation, DM, Duchenne-type muscular dystrophy, and melanomas

82
Q

Globulins

  • A light chain component of the major histocompatibility complex class I (HLA)
  • Found on the surface of most nucleated cells and is present in high concentrations on lymphocytes
  • GFR: freely filtered by the glomerulus but reabsorbed by PCT
  • Is measured by immunoassays
A

β2 - Microglobulin

83
Q

Globulins

Refers to chylomicrons, VLDL, LDL, and HDL

A

Lipoproteins

84
Q

β2 - Microglobulin

High or Low?
Impaired clearance by the kidney or overproduction of the protein in rheumatoid arthritis and SLE

85
Q

Globulin

  • One of the largest proteins
  • Glycoprotein synthesized by the liver
  • An acute phase reactant
  • Between the β and γ globulins
  • Forms a fibrin clot when activated by thrombin
  • Marker for long term prognosis of cardiovascular disease
A

Fibrinogen

86
Q

Fibrinogen

High or Low?
Inflammation, pregnancy, and use of oral contraceptive

87
Q

Fibrinogen

High or Low?
Extensive coagulation

88
Q

Globulins

  • From the liver
  • A natural defense mechanism from infection
  • Classical, Lectin, and Alternative pathways (when activated, the membrane attack complex begins to lyse the foreign body)
  • Consumed in inflammatory reaction (decreased)
A

Complement

89
Q

Complement

Elevated or Decreased?
Have little clinical importance

90
Q

Elevated or Decreased?
The complement may be consumed (e.g. in SLE and RA) or it may be decreased (malnutrition) or absent due to genetic defects

91
Q

Complement

What are the 5 membrane attack complexes?

A

C5, C6, C7, C8, and C9

92
Q

Globulins

**- From the liver, it precipitates with the C-polysaccharide (from pneumococci)
- Migrates toward the Beta-region (Bishop)/Gamma-region (Henry’s)
- APR - one of the first to rise in inflammation or tissue necrosis (not specific)
- bound to bacteria facilitates opsonization, initiates coagulation, and complement activation
- Can be used as a screening test to determine if the infection is bacterial or viral
- Binds with phosphocholine and LDL

A

C-Reactive Protein (CRP)

93
Q

C-Reactive Protein (CRP)

High or Low?
Acute infection, inflammatory disease, tissue necrosis, acute rheumatic fever, bacterial infections, myocardial infarction, rheumatoid arthritis, carcinomatosis, gout, and viral infections

94
Q

C-Reactive Protein (CRP)

  • A condition that involves hsCRP
  • Recurrent event in patients with unstable angina and AMI
  • Low survival rate from CVD
A

Atherosclerosis

95
Q

Globulins

  • Migrates toward the gamma-region
  • Are not synthesized by neonates
A

Immunoglobulins

96
Q

Immunoglobulins

  1. Crosses the placenta thus it comes from the mother
  2. Does not cross the placenta (synthesized by the neonate upon reaching 1 year old); these are also higher in females
  3. Generally higher in males (present in secretions)
  4. For parasitic and allergic conditions
  5. No known function

A. IgD
B. IgA
C. IgG
D. IgM
E. IgE

97
Q

Acute Phase Reactants

Familiarize yourself with the APRs

A

APRs:
- Antitrypsin
- Acid Glycoproteins
- Antichymotrypsins
- Thyroxine-binding Globulins
- A2 Macroglobulins
- Ceruloplasmin
- Haptoglobin
- LDL
- Hemopexin
- Complement
- CRP
- Fibrinogen
- Immunoglobulins

98
Q

Patterns of Electrophoresis

Immediate Response Graph:
What regions increase and what decrease?

(2 answers)

A

Increased: A2 region (an APR)
Decreased: Albumin region (a negative APR)

99
Q

Patterns of Electrophoresis

Delayed Response Graph:
What regions increase and what decrease?

(3 answers)

A

Increased: A2 region (haptoglobin: an APR) and Gamma Region (IgG)
Decreased: Albumin

100
Q

Patterns of Electrophoresis

Hypogammaglobulinemia Graph:
What region decreases?

A

Gamma region (gammaglobulins in the blood)

Occurs in neonates (not yet capable of producing antibodies), undergoing chemotherapy, immunocompromised, lymphoreticular disorders, or agammaglobulinemia (total absence)

101
Q

Patterns of Electrophoresis

Hepatic Cirrhosis/Polyclonal Gammopathy Graph:
What regions increase and decrease?

(2 answers)

A

Increased: Gamma region (beta-gamma bridging); all immunoglobulins are INCREASED
Decreased: Albumin region (the liver produces albumin)

102
Q

Patterns of Electrophoresis

Multiple Myeloma/Paraprotein/Monoclonal Gamopathy Graph:
What regions increase and decrease?

(2 answers)

A

Increased: Only specific immunoglobulins (variable for multiple myeloma)
Decreased: Albumin region

Multiple myeloma: Elevated B-cells/plasma cells

103
Q

Patterns of Electrophoresis

In macroglobulinemia, what immunoglobulin is elevated?

104
Q

Patterns of Electrophoresis

Nephrotic Syndrome Graph:
What regions increase and decrease IN BLOOD?

(3 answers)

A

Increased: A2 Macroglobulins (large protein)
Decreased: Albumin and other regions

Decreased because kidney is damaged therefore these analytes are excreted

105
Q

Patterns of Electrophoresis

Nephrotic Syndrome Graph:
What regions increase and decrease IN URINE?

A

Increased: Albumin and other regions
Decreased: A2 Macroglobulins (large protein)

Reverse of nephrotic syndrome blood levels

106
Q

Patterns of Electrophoresis

Protein-losing Enteropathy Graph:
What regions increase and decrease?

(4 answers)

Protein-losing = absorption problem

A

Increased: Haptoglobin and A2 Macroglobulin
Decreased: Albumin and Gamma-region

Clinical Chemistry (CC 1) (33 decks)

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