Term
| What is the typical structure of an enzyme? |
|
Definition
| Tertiary, globular protein |
|
|
Term
T or F:
Enzymes for side products |
|
Definition
|
|
Term
|
Definition
| inactive form of an enzyme. also called zymogen |
|
|
Term
|
Definition
apoenzyme - w/o cofactor
holoenzyme - w/ cofactor |
|
|
Term
T or F:
Cofactors are protein molecules which assist enzyme function. |
|
Definition
false
not proteins but everything else is true |
|
|
Term
|
Definition
oxidoreductases
transferases
hydrolases
lyases - cleave by means other than oxidation or hydrolysis
isomerase
ligase or synthetases |
|
|
Term
| noncompetitive vs uncompetitive inhibition |
|
Definition
non - inhibitor binds to site other than active site and changes enzyme active site conformation
un - enzyme and substrate bind but inhibitor binds afterward preventing a release |
|
|
Term
| what are we measuring with zeroth order rxn and why? |
|
Definition
| measuring enzyme capability and not the substrate because the [s] does not affect R.O.R. |
|
|
Term
| What type of inhibition uses sulfhydryl groups and heavy metals? |
|
Definition
| Covalent bonding inhibition |
|
|
Term
|
Definition
|
|
Term
|
Definition
|
|
Term
| Relationship between acidity and Ka? |
|
Definition
|
|
Term
| How to calculation H2O dissociation constant (k)? |
|
Definition
|
|
Term
| Definition and function of buffer system> |
|
Definition
weak acid, conjugate base system
prevents dramatic pH change |
|
|
Term
| when is a buffer most effective? |
|
Definition
| When the buffer acid/base ratio is 10:1 or 1:10 but 1:1 is the best |
|
|
Term
| Function of physiological buffers? |
|
Definition
| To maintain blood pH as cell metabolizes waste products |
|
|
Term
| Function of phosphate buffer? |
|
Definition
| increases [] of phosphate in intracellular fluid |
|
|
Term
| Henderson hasselbach equation |
|
Definition
|
|
Term
| start and end products of anaerobic glycolysis? |
|
Definition
| transformation of glucose to pyruvate |
|
|
Term
| Where does 1st ATP molecule come from during anaerobic glycolysis? |
|
Definition
| 1,3-BPG becoming 3-phosphoglycerate |
|
|
Term
| Where does 2nd ATP molecule come from during anaerobic glycolysis? |
|
Definition
|
|
Term
| Rate limiting enzyme of anaerobic glycolysis? |
|
Definition
|
|
Term
| What is needed in order for hemoglobin to release oxygen |
|
Definition
|
|
Term
T or F:
OAA helps jumpstart gluconeogenesis |
|
Definition
|
|
Term
| How many ATP are produced from TCA? |
|
Definition
|
|
Term
| What enzyme is needed for glycogenesis? |
|
Definition
|
|
Term
| What does F-1,6-BP become durng anaerobic glycolysis? |
|
Definition
| 2 3 carbon molecules being glyceraldehyde-3-phosphate and dihydroxyacetone phosphate |
|
|
Term
| What enzyme is needed in order for 1,3-BPG to become 3-phosphoglycerate? |
|
Definition
|
|
Term
| What enzyme helps convert g3p to 1,3-bpg? |
|
Definition
|
|
Term
| Which anaerobic glycolysis intermediate is used for the pentose phosphate shunt? |
|
Definition
|
|
Term
| primary product of pentose phosphate shunt? how is product generated |
|
Definition
| NADPH from NADP via oxidoreductase enzymes |
|
|
Term
| Importance of pentose phosphate shunt |
|
Definition
| preventing oxidative damage to RBC's |
|
|
Term
| 2,3-BPG pathway initiating molecule |
|
Definition
|
|
Term
| Location of TCA cycle? function |
|
Definition
mitochondria
strips electrons from free molecules |
|
|
Term
|
Definition
|
|
Term
| starting and ending molecule of TCA cycle? |
|
Definition
Start- Acetyl CoA
End- OAA |
|
|
Term
| What two ways can acetyl coA be formed? |
|
Definition
1) directly from fatty acid metabolism
2) form pyruvate at end of protein and carb metabolism |
|
|
Term
| How does gluconeogenesis initiate? |
|
Definition
| By hormones when body is in fasting state |
|
|
Term
| Start and end products of gluconeogenesis? |
|
Definition
Start- pyruvate
End - G-6-P (NOT glucose) glucose conversion takes places in the smooth ER and then released into cytoplasm |
|
|
Term
| Which metabolic process helps maintain homeostasis of glucose in the body? |
|
Definition
|
|
Term
| Increasing levels of cAMP, glucagon, and epinephrine will stimulate this metabolic process? |
|
Definition
|
|
Term
| Glycogenolysis is regulated by ... |
|
Definition
|
|
Term
|
Definition
| 3 carbon chain with OH on each |
|
|
Term
| What category of lipid is a wax |
|
Definition
|
|
Term
|
Definition
| long carbon chains with carboxyl group on end |
|
|
Term
| A neutral fat contains... |
|
Definition
| ester of fatty acid, glycerol, free fatty acid chain |
|
|
Term
| What makes up a lecithin? How does this compare to cephalin? |
|
Definition
glycerol, fatty acid, phosphoric acid residue, choline
cephalins contain everything except choline |
|
|
Term
|
Definition
| Molecules derived from hydrolysis of simple and conjugated fats |
|
|
Term
| Isoprenoids are... and are used to synthesize... |
|
Definition
| repeating 5C structural units synthesized from acetyl coa and are used to synthesize terpenes and steroids |
|
|
Term
| Which lipoprotein does the liver synthesize? |
|
Definition
|
|
Term
| HDL's highest % substituent is... and function to... |
|
Definition
| phospholipid functions to move cholesterol of the liver |
|
|
Term
| LDL's highest % substituent is... and comprises of apoprotein... |
|
Definition
|
|
Term
| Chylomicrons have the highest %... |
|
Definition
|
|
Term
| lipid synthesis requires what molecule to start? Cholesterol synthesis? |
|
Definition
|
|
Term
| What is the significant enzyme for lipid synthesis? |
|
Definition
|
|
Term
| What is the end product of lipid synthesis? |
|
Definition
|
|
Term
| Acetyl-CoA becomes what during cholesterol synthesis? |
|
Definition
|
|
Term
| The rate limiting enzyme during cholesterol synthesis is... |
|
Definition
|
|
Term
| HMG-CoA becomes _______ during cholesterol synthesis? |
|
Definition
|
|
Term
| What is the precursor to the activated isoprenoid during cholesterol synthesis? |
|
Definition
|
|
Term
| The precursor to cholesterol is... |
|
Definition
|
|
Term
| The 4 steps of lipid catabolism are... |
|
Definition
| Dehydrogenation, hydration, oxidation, hydrolysis |
|
|
Term
| What is the product of step 1 during lipid catabolism? |
|
Definition
|
|
Term
| Lipid catabolism dehydrogenation occurs in the... |
|
Definition
| ER and outer mitochondrial membrane |
|
|
Term
| What happens to enoyl CoA during Lipid catabolism? |
|
Definition
| gets hydrated to form 3-hydroxyacyl coa |
|
|
Term
| 3-hydroxyacyl coa becomes ________ during lipid catabolism |
|
Definition
|
|
Term
| What are the final products of fatty acid catabolism (general) |
|
Definition
|
|
Term
| HMG CoA gets formed by the combination of acetyl coa and __________________ |
|
Definition
|
|
Term
| B-oxidation creates a DB by _____________________ which can be ________ so that 2 more carbons can be chopped off |
|
Definition
removes H+ from B-carbon to form DB
gets hydrated |
|
|
Term
| How many atps get formed during lipid catabolism? |
|
Definition
|
|
Term
| type of bond linking a.a.'s |
|
Definition
|
|
Term
| What causes chain folding of the secondary structure (proteins)? |
|
Definition
|
|
Term
| What is the H+ donor and acceptor of the secondary structure (Proteins)? |
|
Definition
| amine group donor, carbonyl group acceptor |
|
|
Term
| Describe the bonding of an a-helix? |
|
Definition
| h-bond forms from amine group and carbonyl group 4 residues away |
|
|
Term
| describe the bonding of a b-sheet? |
|
Definition
| h-bond forms from adjacent amine and carbonyl groups |
|
|
Term
| The core of a tertiary structure is (hydrophobic/hydrophilic) while the surface is (hydrophobic/hydrophilic). |
|
Definition
Core - hydrophobic
Surface - Hydrophilic |
|
|
Term
T or F:
Hydrophilic interactions are major driving force for protein folding. |
|
Definition
False
Hydrophobic interactions |
|
|
Term
|
Definition
|
|
Term
|
Definition
|
|
Term
| A cavity of a globular protein is a... |
|
Definition
| waterless region where a ligand binds |
|
|
Term
| What type of protein structure does "covalent-crosslinking" occur in |
|
Definition
|
|
Term
| How do heavy metals denature proteins? |
|
Definition
| disrupt salt bridges and bind sulfhydryl groups |
|
|
Term
| How are strong acids/bases denaturing groups? |
|
Definition
| disrupt h-bonding and salt bridges |
|
|
Term
T or F:
Solvents/detergents denature proteins by disrupting hydrophilic interactions |
|
Definition
F
hydrophobic interactions |
|
|
Term
| What are examples of eicosinoids? |
|
Definition
| prostaglandin, thromboxane, leukotriene |
|
|
Term
| What classifies a peptide hormone? Where are they secreted from? |
|
Definition
| 50 or less A.A.'s produced by A.P., hypothalamus, A. medulla, nervous tissue |
|
|
Term
| What second messenger activates protein kinase |
|
Definition
|
|
Term
| What is significant about the phosphoinositide signaling system? |
|
Definition
| allows Ca++ to move into cytosol |
|
|
Term
| Where are steroid hormones secreted from |
|
Definition
| adrenal cortex and gonads |
|
|
Term
|
Definition
| base and 5C sugar without phosphate group |
|
|
Term
| Nucleotides are linked by what type of bonds? |
|
Definition
|
|
Term
T or F
5' OH groups and 3' Phosphate groups bind in DNA formation |
|
Definition
|
|
Term
| What is the charge of the exposed phosphate group of DNA |
|
Definition
|
|
Term
| What direction does DNA polymerase read? |
|
Definition
|
|
Term
| What type of molecule initiates DNA replication? |
|
Definition
|
|
Term
| DNA is replicated in what direction? |
|
Definition
|
|
Term
| How does DNA elongation occur? |
|
Definition
| H-bonding of nucleotides to nitrogenous bases |
|
|
Term
| Which DNA polymerase is used on the leading strand? Direction? |
|
Definition
|
|
Term
| Which DNA polymerase is used on the lagging strand? Direction? |
|
Definition
DNA pol. I and III
I- replaces RNA primers
III- fills in new DNA
5' --> 3' |
|
|
Term
|
Definition
| recognizes nicks and removes mismatched pairs. |
|
|
Term
| What part of the cell does transcription occur? |
|
Definition
|
|
Term
|
Definition
| dna wound around histones |
|
|
Term
| which RNA pol binds to sites of tandem repeats? |
|
Definition
|
|
Term
|
Definition
| TATA box or CAAT box, downstream of start site |
|
|
Term
| RNA Pol III's promoter region is located... |
|
Definition
| in the transcribed sequence |
|
|
Term
| Most rRNA is synthesized from which polymerase? mRNA? tRNA? |
|
Definition
rRNA - pol I
mRNA - pol II
tRNA - pol III |
|
|
Term
| When does mRNA become functional> |
|
Definition
|
|
Term
| major product of transcription? |
|
Definition
|
|
Term
| function of aminoacyl tRNA synthetase? |
|
Definition
| activates linkage of specific AA to appropriate tRNA |
|
|
Term
| function of peptidyl transferase? |
|
Definition
allows formation of peptide bonds between a.a.'s
also cleaves once STOP is reached |
|
|
Term
|
Definition
mis- changes aa sequence
non - changes to stop codon |
|
|
Term
| Use of southern blotting? Northern? Western? |
|
Definition
Southern - DNA
Northern - RNA
Western - protein |
|
|
Term
| 5 steps of southern blotting? |
|
Definition
1) R.E. cuts DNA
2) electrophoresis
3) depurination, denaturation
4) transfer to solid support
5) probe added |
|
|
Term
| What chemical is used for depurination? What type of fragement is best suited? |
|
Definition
used for 500 BP or greater frags
gel exposed to HCl |
|
|
Term
| What chemical denatures with southern blotting? |
|
Definition
|
|
Term
| What must occur to nitrocellulose membrane in order for it to be of use? |
|
Definition
| moistened with buffer solutin |
|
|
Term
| w/ organic extraction, how is DNA extracted? |
|
Definition
| by phenol-chloroform mixture which removes protein material |
|
|
Term
| W/ organic extraction, how is dna ppt'd? |
|
Definition
| by addition of ice cole EtOH and salt |
|
|
Term
| w/ inorganic extraction, how are proteins extracteed? |
|
Definition
| By addition of LiCl and centrifugation which pellets them |
|
|
Term
| How is dna ppt'd with inorganic extraction? |
|
Definition
|
|
Term
| Function of detergents with dna extraction? |
|
Definition
|
|
Term
| If you change [ ] of agar, what occurs? |
|
Definition
Size of the pores change (gets smaller)
inversely proportional |
|
|
Term
|
Definition
|
|
Term
|
Definition
sits in minor groove of double helix
much more [ ]'d than EtBr |
|
|
Term
| Silver stain is used... and is good for |
|
Definition
post electrophoresis
good for protein analysis |
|
|
Term
| What type of groups does silver stain react with? |
|
Definition
|
|
Term
| Silver stain samples are fixed with _____ and _____ |
|
Definition
|
|
Term
|
Definition
|
|
Term
| how is pcr denaturing accomplished? |
|
Definition
| brief heating to break H-bonds |
|
|
Term
| how is pcr annealing accomplished? |
|
Definition
| primers form h-bonds with target sequence during a brief cooling period |
|
|
Term
| how does primer extension occur during pcr? |
|
Definition
| dna polymerase adds nucleotides to the 3/ end of each primer |
|
|
Term
| The pcr master mix consists of... |
|
Definition
|
|
Term
| temp range for denaturation? annealing? extension? |
|
Definition
denaturing - 90-96 deg C
annealing - 50-70 deg C
extension - 68-75 deg C |
|
|
Term
|
Definition
| more than 1 primer in mix allowing multiple region copying |
|
|
Term
|
Definition
| adds label to mix so amplicons show results as the test is running |
|
|
Term
|
Definition
| 2 sets of primers used in sequence increasing specificity and sensitivity |
|
|
Term
| reverse transcriptase pcr |
|
Definition
used when using rna sample
forms cDNA which is then used for PCR |
|
|
Term
|
Definition
10-50 bases repeat in tandem
detected by southern blot and pcr |
|
|
Term
|
Definition
1-10 bases repeat in tandem
detected by pcr |
|
|
Term
single strand conformation polymorphisms
(what forms what? what happens to the product?) |
|
Definition
| denature dilute dsDNA, resulting ssDNA will form 3-d structure which is separated by gel electrophoresis |
|
|
Term
Denaturing gradient cells
(process, what to do w/ results) |
|
Definition
| denature dsDNA as it moves down different [ ]'s of agar. Then compare w/ known results to find mutation and polymorphisms in populations |
|
|
Term
| Melting curve analysis (what results mean, how to make results more efficient) |
|
Definition
delta Tm means a mutation is present
enhanced by FRET |
|
|
Term
| What 4 items are required in order to run genetic or parental testing? |
|
Definition
| consent, transfusion history, photo id, ethnicity |
|
|
Term
| What type of blood tube is used for HIV HCV patients? |
|
Definition
|
|
Term
| How can a lab validate in-house testing method? |
|
Definition
| sensitivity, specificity, accuracy |
|
|
Term
| How to properly clean work area? |
|
Definition
|
|
Term
| What are internal controls necessary? |
|
Definition
| to detect target specific controls |
|
|
Term
| Std. curves mark changes in _______ |
|
Definition
|
|
Term
| W/ PCR, what does a pos. control assure? |
|
Definition
|
|
Term
| w/ PCR what does an amp. control assure? |
|
Definition
| thermocycler and reagents are working under conditions necessary for target |
|
|
Term
| w/ PCR what does a neg. control assure? |
|
Definition
| no contamination is present |
|
|
Term
| W/ PCR what does a neg. template control assure? |
|
Definition
| primers are not annealing errantly |
|
|
Term
| What techniques prevent contaminants from mispriming? |
|
Definition
|
|
Term
| How often are incubators monitored? |
|
Definition
|
|
Term
| How often are pipettes checked? |
|
Definition
|
|
Term
| centrifuges are checked... |
|
Definition
|
|
Term
|
Definition
| clinical lab improvement amendments of 1988 |
|
|
Term
|
Definition
| CLS institute- establishes acceptable methods and practices |
|
|
Term
|
Definition
| College of American pathologists- require proficiency testing |
|
|
Term
|
Definition
| health info portability and accessibility act |
|
|
Term
| IHC tests are used to.... while FISH tests are used to... |
|
Definition
|
|
Term
| what test is used to confirm sickel cell anemia? |
|
Definition
|
|
Term
| Factor V leiden can be confirmed by performing... |
|
Definition
|
|
Term
| Cystic Fibrosis is confirmed by... |
|
Definition
| RFLP, PCR-RFLP, ss conform polymorphism |
|
|
