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53 Cards in this Set
- Front
- Back
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List the 8 functions of proteins:
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1. Structural
2. Hormonal 3. Catalytic (enzymes) 4. Storage/transport 5. Immunity 6. Receptors 7. Osmotic balance 8. Nutritional/maintenance |
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What characterizes primary structure?
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-Linear amino acid sequence
-Disulfide bridges |
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What characterizes 2ndary structure?
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Shape of chains - alpha helix or beta sheets
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What characterizes tertiary structure?
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The 3-D shape of entire chain; determines protein function.
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What characterizes quaternary structure?
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2 or more polypeptide chains held together noncovalently.
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Which level of structure is disruptured by denaturation?
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Tertiary
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What are the components of a protein electrophoresis?
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-Prealbumin
-Albumin -Alpha-1 -Alpha-2 -Beta -Gamma |
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What are the negative APR's?
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-Prealbumin
-Transferrin |
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What are the normal APR's?
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-a-antitrypsin
-Haptoglobin -C-reactive protein -Antithrombin-III |
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which protein is associated with non-environmental emphysema?
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Antitrypsin
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Which protein is increased in lupus and rheumatoid arthritis?
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Acid glycoprotein
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What makes up the alpha-1 fraction of electrophoresis?
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-Antitrypsin
-Acid glycoprotein |
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What makes up the alpha-2 fraction?
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-Ceruloplasmin
-Macroglobulin -Haptoglobin |
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What makes up the Beta-1 fraction?
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-Hemopexin
-Transferrin |
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What makes up the Beta-2 fraction?
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-Complement (CRP)
-Fibrinogen -AT-III |
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What is Wilson's disease?
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An inability to process copper correctly
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What is an indicator of Wilson's disease?
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Decreased Ceruloplasmin
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Which APR is significantly elevated in infections?
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C-reactive protein
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Which protein is NOT lost in renal damage b/c of its large size?
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Alpha2-macroglobulin
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What is the historical method for protein measurement?
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Kjeldahl
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What is the principle of the Kjeldahl method?
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-Protein is digested w/ acid
-Nitrogen converted to NH4+ -Protein nitrogen expressed as total protein by 6.25 factor |
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What is the current precipitation method for protein called?
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Turbidimetric
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What is used for the turbidimetric method?
How is it done? |
Sulfasalicylic acid - precipitates protein for scattered light measurement.
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What is used to measure the light scatter in the turbidimetric method?
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Nephelometer
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What is the principle of refractometry?
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Proteins change refractive index of water in propertion to their concentration.
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What are 2 drawbacks of refractometry?
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-Only gives an estimate of protein concentration.
-Interferences = high BUN, hyperglycemic, lipemic, elevated albumin levels. |
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What is the principle of the Biuret method?
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-Protein + Cu2+
-Alk pH via sodium tartrate -Violet color forms by chelation of Cu and peptide bond |
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What wavelength is the biuret method read at?
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540 nm
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What are drawbacks of the biuret method?
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-Hemolysis interferes
-Gross lipemia interferes -Not useful for CSF/urine |
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What method is used for Albumin analysis?
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-Dyebinding method
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-What dye is used for Albumin?
-What needs to be kept in mind? |
Brom-cresol Green
-Read at spcf time before the dye binds other proteins. |
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What protein method is useful for CSF and urine?
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Immunochemical methods
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How are immunochemical methods read for protein?
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-Nephelometry
-Turbidometric -RIA |
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What charge is on the
-Cathode -Anode |
Cathode = negatively charged
Anode = positively charged |
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So what migrates to the
-Cathode -Anode |
Cations migrate to the cathode
Anions migrate to the anode. |
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What factors affect mobility in serum protein electrophoresis?
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-Net protein charge
-Protein size, shape -Electrical field -Support used (gel, etc) -Temperature -Buffer |
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How does protein size affect electrophoresis?
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-Larger proteins migrate slower
-Smaller proteins migrate faster |
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What is the definition of electrophoresis?
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-Differential migration of charged particles in a liquid medium under an electrical influence causing them to seperate.
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What type of medium is typically used for protein electrophoresis?
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Cellulose acetate
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What is the principle of isoelectric focusing?
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-A pH gradient is set up on the solid medium
-Proteins migrate to their pI -Allows specific seperation |
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How are urine samples prepped for electrophoresis?
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Need to be concentrated
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What things are looked for in urine protein electrophoresis?
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-Bence jones proteins
-Albumin |
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Bence jones proteins in urine are an indicator of:
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Multiple myeloma
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What type of CSF has the most protein normally?
-Ventricular -Cisternal -Lumbar |
Ventric < Cistern < Lumbar
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What are increased CSF proteins an indicator of?
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Meningitis or BBB integrity damage
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How is radial immunodiffusion used for protein measurement?
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The area of diffusion in agar is specific for proteins, and allows quantitation of the concentration.
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How is Ouchterlony analysis useful in protein analysis?
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Allows for determining identity, partial identity, or non-identity of proteins.
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How is Rocket electroimmunoassay analysed?
What type of protein is this typically done for? |
By measuring the height of the precipitation the ANTIGEN concentration is measured.
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What is "electro-endosmosis"?
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A net flow of solvent to the cathode - an interferent
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How does electroendosmosis interfere with protein electrophoresis?
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It hinders solutes that want to go to the anode.
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How does HEAT affect electrophoresis?
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-Increases migration
-distorts patterns -Denatures some proteins -Increases ionic concentration of the buffer |
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What type of buffer enhances electrophoresis?
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Buffers with a high ion cloud
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What is a densitometer?
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modified spec for quantitating seperated bands
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