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Clinical Chemistry > Total Protein > Flashcards

Total Protein Flashcards

1
Q

Protein

A
  • constituents include C, N, H and O
  • peptide bonds are between amino (NH2) and caboxyl (C=0) groups
  • amino acids are zwitterions (ampholytic)
  • 15-16% nitrogen
  • coded by DNA, made via transcription and translation
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2
Q

Protein structure

A
  • primary: chain of AA’s
  • secondary: pleated sheets and helices
    tertiary: folded up form
  • quaternary: multiple tertiary structures folded together
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3
Q

Aminoacidopathies

A
  • PKU
  • Tyrosinemia
  • Alkaptonuria
  • Maple Syrup Disease
  • Isovaleric Acidemia
  • Homocystinuria
  • Citrullinemia
  • Arginiosuccinic Aciduria
  • Cystinuria
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4
Q

PKU

A
  • lack of phenylalanine hydroxylase
  • leads to build up of phenylalanine in the blood
  • excess phenylalanine can lead to mental retardation
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5
Q

Tyrosinemia

A
  • 3 types
  • Type I: fumarylacetoacetate hydrolase deficeincy
  • Type II: tyrosine aminotransferase deficiency
  • Type III: 4-hydroxyphenylpyruvate dioxygenase
  • Type II is main one seen
  • Type III is rare
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6
Q

Alkaptonuria

A
  • lack of homogentisate oxidase

- accumulation of homogentisic acid turns urine black

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7
Q

Maple Syrup Disease (MSUD)

A
  • decreased or absent branched-chain alpha-ketoacid decarboxylase
  • build up of branched chain amino acids smell like maple syrup
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8
Q

Isovaleric Acidemia

A

Isovaleryl-Co-A dehydrogenase deficiency

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9
Q

Homocystinuria

A

cystathionine-beta-synthetase deficiency

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10
Q

Citrullinemia

A

Type I: argininosuccinic acid synthetase

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11
Q

Arginiosuccinic Aciduria

A

arginosuccinic acid lyase deficiency

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12
Q

Cystinuria

A
  • defect in amino acid transport system
  • kidneys don’t filter cysteine properly
  • higher levels of cysteine in the urine and blood
  • precipitates can form stones
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13
Q

Protein involvement

A
  • muscle movement
  • enzymes
  • structure of cells
  • extracellular matrix (collagen)
  • transportation of molecules
  • antibodies
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14
Q

Protein functions

A
  • energy
  • osmotic force
  • acid-base balance
  • transport
  • antibodies
  • hormones
  • enzymes
  • hemostasis
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15
Q

Two major groups of protein in plasma

A

albumin and globulin

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16
Q

Other tissue proteins

A

collagen, elastin, keratin

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17
Q

Conjugated proteins

A

nucleoproteins, glycoproteins, lipoproteins

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18
Q

Albumin

A
  • most abundant serum protein
  • involved in transport of other molecules
  • involved in osmotic balance
  • made in liver with 585 AA’s (small)
  • buffers pH
  • negative acute phase reactant (concentration decreases in acute phase reaction)
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19
Q

Globulin

A
  • many specific proteins
  • antibodies are included in this group
  • alpha, beta and gamma bands on electrophoresis
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20
Q

Pre-albumin (Transthyretin)

A
  • transport protein for T4 and T3
  • binds retinol-binding protein to transport Vitamin A
  • decreased when decreased protein synthesis or liver disease
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21
Q

Molecules albumin binds/transports

A
  • T4 and T3
  • fat-soluble hormones, iron, fatty acids
  • unconjugated bilirubin
  • calcium, magnesium, ions
  • drugs, aspirin
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22
Q

Causes of Albumin decrease

A
  • liver disease
  • protein loss: GI inflammation, diarrhea
  • kidney disease, nephrotic syndrom, burns, IV fluid excess
  • acute disease, mutations, redistribution from hemodilution
  • increased capillary permeability, decreased capillary clearance
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23
Q

alpha-1 globulins

A
  • alpha-1-Antitrypsin (AAT)
  • alpha-1-Fetoprotein (AFP)
  • alpha-1-Acid Glycoprotein or Orosomucoid
  • alpha-1-Antichymotrypsin
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24
Q

alpha-1-Antitrypsin (AAT)

A
  • synthesized in the liver
  • inhibits neutrophil elastase
  • increased in acute phase reaction, pregnancy and contraception meds
25
Q

alpha-1-Fetoprotein (AFP)

A
  • synthesized in developing embryo, then parenchymal cells of liver
  • decreased concentration 8-12 months after birth
  • binds to estradiol
  • marker of spina bifida and neural tube defects in fetus
  • low AFP in mom can indicate Down’s syndrome
26
Q

alpha-1-Acid Glycoprotein (Orosomucoid)

A
  • synthesized in the liver
  • acute phase reactant
  • similar structure to Ig’s
  • elevated in stress, trauma, AMI, inflammation, pregnancy, cancer, pneumonia, RA, surgery
27
Q

alpha-1-Antichymotrypsin

A
  • synthesized in the liver
  • inhibits enzymes of cathepsin G, pancreatic elastase, mast cell chymase, and chymotrypsin
  • unchecked these would eat away at GI tissue
  • acute phase reactant
28
Q

alpha-2 globulins

A
  • Gc-globulin
  • Haptoglobin
  • Ceruloplasmin
  • alpha-2-Macroglobulin
29
Q

Gc-globulin

A
  • Vitamin D-binding protein
  • synthesized in liver hepatocytes
  • carries Vitamin D and fatty acids
30
Q

Haptoglobin

A
  • synthesized in the liver
  • acute phase reactant
  • binds hemoglobin
  • increases in RA, burns, nephrotic syndrome and Type 2 DM
  • decreased in hemolytic anemia (when Hb is decreased)
31
Q

Ceruloplasmin

A
  • synthesized in the liver
  • acute phase reactant
  • has 8 Cu’s attached
  • increases in inflammation, severe infections, cancer and pregnancy
  • decreases in malnutrition, malabsorption, liver disease, nephrotic syndrome and Menkes syndrome
32
Q

alpha-2-Macroglobulin

A
  • synthesized in the liver
  • major component of alpha-2 band
  • inhibits trypsin, thrombin, kallikrein and plasmin
  • increased in DM, liver disease, nephrosis, pregnancy and BC pill
33
Q

Beta globulins

A
  • transferrin
  • hemopexin
  • lipoproteins
  • beta-2-Microglobulin
  • Complement
34
Q

Transferrin

A
  • synthesized in the liver
  • negative acute phase reactant
  • transports iron (Fe3+)
  • increased in anemia, hemochromatosis
  • decreased in malnutrition, liver disease, kidney loss
35
Q

Hemopexin

A
  • synthesized in the liver
  • acute phase reactant
  • takes heme back to liver for degradation and iron recovery
  • increased in inflammation, diabetes, Duchenne muscular dystrophy and melanomas
  • decreased in hemolytic anemia
36
Q

beta-2-Microglobulin

A
  • small concentrations present

- surface of nucleated cells and lymphocytes

37
Q

Complement

A

C3 and C4 increase in inflammation

38
Q

Proteins between beta and gamma

A
  • Fibrinogen

- C-Reactive protein

39
Q

C-Reactive Protein

A
  • acute phase reactant
  • rises sharply with inflammation
  • increases in Atherosclerosis, Cardiovascular disease and Diabetes
  • hs-CRP is marker for cardiovascular disease
40
Q

Gamma globulins

A
  • Immunoglobulins (glycoproteins)
  • synthesized in B cells
  • IgG is most abundant
  • individual concentrations determined by immunoassay
41
Q

Other Proteins

A
  • Myoglobin
  • Troponin
  • Brain Natriuretic Peptide (BNP)
  • Cystatin C
  • Amyloid
42
Q

Myoglobin

A
  • contains heme
  • carries O2 to muscle
  • increases 1st in AMI
43
Q

Troponin

A
  • cardiac forms of cTnT and cTnI used to detect AMI

- better than CK-MB

44
Q

Brain Natriuretic Peptide (BNP)

A
  • marker for Congestive Heart Failure
  • found in left ventricular myocytes
  • released when fluid overload to get kidneys to excrete fluid
  • BNP = active
  • N-terminal BNP = inactive
45
Q

Cystatin C

A
  • cysteine proteinase inhibitor
  • used to monitor GFR
  • not affected by factors that alter creatinine excretion
  • increases when GFR decreases
46
Q

Amyloid

A
  • insoluble fibrous protein aggregate when beta-pleated sheets are altered
  • deposits can lead to organ failure
  • seen in chronic infections, malignancies, rheumatoid disorders
  • low Amyloid beta42 and high Tau = likelihood of Alzheimers
  • high Amyloid beta42 and low Tau = dementia
47
Q

Total protein

A
  • sum of all albumin and globulins
  • readings in grams/dL
  • reflects nutritional status, kidney function, liver function, etc.
  • change in total protein reflects change in nitrogen status and non-protein nitrogen
48
Q

Hyperproteinemia

A
  • increase in total protein
  • may not be due to disease
  • nitrogen content is variable
  • increases with dehydration, excess synthesis (mainly gamma globulins)
49
Q

Hypoproteinemia

A
  • decrease in total protein
  • primarily loss of albumin
  • negative nitrogen balance
  • decreases due to protein loss (kidney, GI, blood loss, burns), lack of protein intake (malnutrition, malabsorption), liver disease (decreased synthesis), immunodeficiency, increased breakdown of proteins (burns, trauma, injury)
50
Q

Protein Methods

A
  • Kjeldahl method
  • Biuret method
  • Refractory method
  • Electrophoresis
  • Nephlometry
  • Colored dyes (albumin)
51
Q

Kjeldahl method

A
  • classic method

- breaks up protein and measures total amount of nitrogen

52
Q

Biuret Reaction

A
  • copper ion binds to at least 2 peptide bonds (chelation)
  • complex is colored and read by spectrophotometer
  • Na-K tartrate complexes with cupric ions to prevent ppt in alkaline pH
  • K-iodine is antioxidant to keep Cu in +2 state (needed for binding)
  • INTERFERENCE: Lipemia
53
Q

Refractory Measurement

A
  • protein in solution (urine) will refract light read by refractometer
  • not as accurate as other methods
  • INTERFERENCES: elevated solutes of glucose, urea, bilirubin and lipids
54
Q

Electrophoresis

A
  • separation of proteins based on electric charges and densities
  • velocity of movement depends on field strength, size and shape of protein molecules, temperature of the buffer, and characteristics of the buffer (pH, etc.)
  • semi-quantitative
55
Q

Nephlometry

A
  • specific antibodies to a specific protein form Ag-Ab complexes
  • measures light scatter due to complexes
  • can also use fluorescent tags for detection
  • standard curves are used to calculate concentration of specific proteins
56
Q

Urinary protein

A
  • Dipstick protein
  • normally negligible
  • albumin (small molecule)
  • Tamm-Horsfall (protein from tubules)
57
Q

Microalbumin

A

used in diabetics to monitor renal function

58
Q

CSF protein

A
  • blood-brain barrier damage leads to increased protein in CSF
  • CSF electrophoresis used to detect MS
  • normal CSF electrophoresis shows pre-albumin, albumin, alpha-1 (antitrypsin), alpha-2 (haptoglobin and ceruloplasmin), beta (transferrin), and gamma (IgG and IgA)
  • MS shows oligoclonal bands in gamma region

Clinical Chemistry (8 decks)

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