Glossary of 14. Proteins
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- List the 8 functions of proteins:
- 1. Structural
3. Catalytic (enzymes)
7. Osmotic balance
- What characterizes primary structure?
- -Linear amino acid sequence
- What characterizes 2ndary structure?
- Shape of chains - alpha helix or beta sheets
- What characterizes tertiary structure?
- The 3-D shape of entire chain; determines protein function.
- What characterizes quaternary structure?
- 2 or more polypeptide chains held together noncovalently.
- Which level of structure is disruptured by denaturation?
- What are the components of a protein electrophoresis?
- What are the negative APR's?
- What are the normal APR's?
- which protein is associated with non-environmental emphysema?
- Which protein is increased in lupus and rheumatoid arthritis?
- Acid glycoprotein
- What makes up the alpha-1 fraction of electrophoresis?
- What makes up the alpha-2 fraction?
- What makes up the Beta-1 fraction?
- What makes up the Beta-2 fraction?
- -Complement (CRP)
- What is Wilson's disease?
- An inability to process copper correctly
- What is an indicator of Wilson's disease?
- Decreased Ceruloplasmin
- Which APR is significantly elevated in infections?
- C-reactive protein
- Which protein is NOT lost in renal damage b/c of its large size?
- What is the historical method for protein measurement?
- What is the principle of the Kjeldahl method?
- -Protein is digested w/ acid
-Nitrogen converted to NH4+
-Protein nitrogen expressed as total protein by 6.25 factor
- What is the current precipitation method for protein called?
- What is used for the turbidimetric method?
How is it done?
- Sulfasalicylic acid - precipitates protein for scattered light measurement.
- What is used to measure the light scatter in the turbidimetric method?
- What is the principle of refractometry?
- Proteins change refractive index of water in propertion to their concentration.
- What are 2 drawbacks of refractometry?
- -Only gives an estimate of protein concentration.
-Interferences = high BUN, hyperglycemic, lipemic, elevated albumin levels.
- What is the principle of the Biuret method?
- -Protein + Cu2+
-Alk pH via sodium tartrate
-Violet color forms by chelation of Cu and peptide bond
- What wavelength is the biuret method read at?
- 540 nm
- What are drawbacks of the biuret method?
- -Hemolysis interferes
-Gross lipemia interferes
-Not useful for CSF/urine
- What method is used for Albumin analysis?
- -Dyebinding method
- -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.
- What protein method is useful for CSF and urine?
- Immunochemical methods
- How are immunochemical methods read for protein?
- What charge is on the
- Cathode = negatively charged
Anode = positively charged
- So what migrates to the
- Cations migrate to the cathode
Anions migrate to the anode.
- What factors affect mobility in serum protein electrophoresis?
- -Net protein charge
-Protein size, shape
-Support used (gel, etc)
- How does protein size affect electrophoresis?
- -Larger proteins migrate slower
-Smaller proteins migrate faster
- What is the definition of electrophoresis?
- -Differential migration of charged particles in a liquid medium under an electrical influence causing them to seperate.
- What type of medium is typically used for protein electrophoresis?
- Cellulose acetate
- What is the principle of isoelectric focusing?
- -A pH gradient is set up on the solid medium
-Proteins migrate to their pI
-Allows specific seperation
- How are urine samples prepped for electrophoresis?
- Need to be concentrated
- What things are looked for in urine protein electrophoresis?
- -Bence jones proteins
- Bence jones proteins in urine are an indicator of:
- Multiple myeloma
- What type of CSF has the most protein normally?
- Ventric < Cistern < Lumbar
- What are increased CSF proteins an indicator of?
- Meningitis or BBB integrity damage
- How is radial immunodiffusion used for protein measurement?
- The area of diffusion in agar is specific for proteins, and allows quantitation of the concentration.
- How is Ouchterlony analysis useful in protein analysis?
- Allows for determining identity, partial identity, or non-identity of proteins.
- 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.
- What is "electro-endosmosis"?
- A net flow of solvent to the cathode - an interferent
- How does electroendosmosis interfere with protein electrophoresis?
- It hinders solutes that want to go to the anode.
- How does HEAT affect electrophoresis?
- -Increases migration
-Denatures some proteins
-Increases ionic concentration of the buffer
- What type of buffer enhances electrophoresis?
- Buffers with a high ion cloud
- What is a densitometer?
- modified spec for quantitating seperated bands
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