Term
|
Definition
*important because dysfunction leads to disease
*needed to solve treatments |
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|
Term
|
Definition
*how DNA carries genetic info in a sequence (DNA = nuc seq)
*4 in DNA/RNA
*COMPOSED: N base, ribose/deoxy, 1+ phosphate
*E carriers(ATP) when release phosphate group
*E storage and transfer to diff molecule
DNA - scribe-> mRNA -lat-> protein |
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|
Term
|
Definition
PURINE
*adenine, guanine
PYRIMIDINE
*cytosine, uracil/thymine |
|
|
Term
|
Definition
*E transferred by phosphates
*humans recycle appropriate ATP/day
*Starch synthesized by adding glucose from repeat ADP gluc |
|
|
Term
|
Definition
* 5' --> 3'
(phosphate) --> (hydroxy)
*phosphodiester bonds link nucleotide residues |
|
|
Term
|
Definition
1) DNA has = 3s of A,T / G,C
2) Base composition varies between species
-25-75 mol (bact)
-40 mol (mammals) for G+C |
|
|
Term
|
Definition
*2, antiparallel polynucleotide strands form DNA helix
*minor/ major groove- uneven width of helix surf
*H bonds cause Chargaff's Rules |
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|
Term
| Complementary Base Pairing |
|
Definition
*bases of opposite strands interacting by H bond
*3 H bond between G,c
2 between A,T
*break G,C uses more E |
|
|
Term
|
Definition
*DNA contains genetic info in form of genes
*example of transformation of patho/non bacterium |
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Term
|
Definition
*DNA duplication during cell division
*results in identical copies |
|
|
Term
|
Definition
| *1 gene scribed/lated to 1 protein |
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|
Term
| Mutation in nucleotide seq |
|
Definition
| * in 3rd position, less immediate effect b/c same AA acid (can be interchanged) |
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Term
|
Definition
| *triplet code of RNA has e. AA is based on it |
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Term
|
Definition
*ability to determine the nuc seq of a gene/ whole genomes
*gives better understanding of molecular mech that support life
*biotech soln (lrg prod of antimalaria, insulin) |
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Term
|
Definition
1)DNA from humans put into plasmid (can bemod)
2) recomb DNA loop
3)replicated to make exact copy
4)ferm tank
5) mult copies of human insulin
6) purify and bottle |
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Term
|
Definition
*allows to insert piece of DNA for replication
*can be mod |
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Term
|
Definition
*antimalria drug
*time consuming found in field plants
*found alt path = more effective |
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Term
|
Definition
*req sm segments of nuc
*76 residues took 7 years
NOW: >3 mil (3Mbp) nuc in hours |
|
|
Term
|
Definition
CATTLE
*painful to dehorn
*introd mutation for cows w/o horns |
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|
Term
| Types of Nucleotide Sequencing Techniques |
|
Definition
1) FRAGMENTATION OF NUCLEOTIDES USING ENZ
-Gel Electrophoresis
2) CHAIN TERMINATOR
-Sanger
3)AMPLIFICATION OF GENE FRAGMENTS
- PCR |
|
|
Term
| Fragmentation Enz for Nucleotides |
|
Definition
NONSPECIFIC
*breaks phosphodiester between residues
*random choice result in mix of fragments of diff lengths
*EX: snake venom phosphodiesterase
SPECIFIC
*bact enz that recognize and cleave DNA that has a specific nucleotide seq
*cause restriction enz that provide protection against viral infection by destruction of foreign(viral) DNA (b/c not yet methylated)
*2 types: endo/exo nucleases |
|
|
Term
|
Definition
*cleave nucleic acids by removing terminal residues
*recognize seq @end of seq
*last nucletide |
|
|
Term
|
Definition
*cleave nucleic acids w/in nucleotide strand
*recognize seq w/in seq
*cleave at specific recognition sites
* 2 types (Type I, II) |
|
|
Term
| Specific Fragmentation of Nucleotides w/ Enz |
|
Definition
1) introduce into bact
2) viruses attach to wall, inject DNA
3) DNA replicate
4) Restriction enz detect / destroy foreign DNA
5)able to determine self/ foreign |
|
|
Term
| Type I & II Endonucleases |
|
Definition
TYPE I
*recognize DNA in specific spot
TYPE II
*recognize a seq close to the potential modification side
*4-8BP; very useful
*recognize palindrome |
|
|
Term
| Understanding Restriction Endonuc Tables |
|
Definition
↓ : cleavage site
Enz Name: abbreviation of microbe in which enz derived from
R: purine
Y: pyrimidine |
|
|
Term
|
Definition
STICKY
*cuts after 1st nucleotide
*self annealing w/ comp base pairs forms H bonds
BLUNT
*cute random/ or in half
*no true, free nucleotide so cant find complementary BP ad cannot self anneal |
|
|
Term
|
Definition
*separate DNA frags by size
*order strands by size
*fragments move toward anode b/c - charge
* - charge on DNA phosp cause move toward + (anode) |
|
|
Term
|
Definition
1) DNA template (1 strand)
2) stepwise add of complementary nuc
-add 5-3
3) add primer
-should be complementary to temp
4) dNTPs pair w/ bases
5) incoming nuc joined to polynuc strand in 5-3
6) catalyzed by DNAP1
7) ddNTP bind to terminate |
|
|
Term
|
Definition
*synthesized short fragments of DNA that is complementary to piece want to amplify
*inc H bond breaks = more E needded to separate fragments |
|
|
Term
|
Definition
*Extends the primer
*uses the free 3'OH to grow strand
*release pyrophosphate (PPi) w/ e. nuc addn
*connect nuc base to form complementary strand (5'->3') |
|
|
Term
|
Definition
*DNA template
*DNAP1
*suitable primer
*4 dNTP substrates w/ free nuc residues (for polymerization)
*sm amount of 2,3-ddNTP
*chromophore (flourescent tag, tells end) |
|
|
Term
|
Definition
*free hyroxyl group on 3' missing so can't cont chain & sequence stops
*sm amount allows rxn to proceed & terminate 1x to amplify seq |
|
|
Term
|
Definition
*polymerase chain rxn
*amplifies gene fragments
*don't want to interrupt rxn
1) HEAT, separate strands (95C)
2) COOL, primer aneealing
-temp: 5->3 ; primer: 3->5
3)add dNTPs & DNAP extend primer
-DNAP survive repeated heat
4) get 2 copies so repeat (25-30x)
5) RESULT: specific fragment of foreign DNA
6) put into recombinant protein
*need mult copies for experiments |
|
|
Term
|
Definition
*DNA temp
*primer
*nucleotides
*polymerase |
|
|
Term
|
Definition
*expression of foreign genes using circular DNA
*foreign DNA obtained by PCR, DNA restriction, chem synth
1) take foreign DNA from PCR want to amplify
2)splice plasmid in seq that has same endonuc
2) put back into bact (pUC18, e.coli)
3) sticky ends anneal w/ ligase (foreign temp glued w/ bact DNA)
3) vector w/ new DNA able to replicate
4) ID cells w/ desired DNA (color change) |
|
|
Term
ID desired cells in Recombinant Proteins
(detecting foreign DNA) |
|
Definition
1) add antibiotics/ chemogenic substances
2) find color change
BLUE: no foreign DNA added
* lacZ encodes for (->) B-galactosidase which cleaves colorless compound X-gal
COLORLESS: foreign DNA
*disrupted LacZ seq and cant -> X-Gal
*resistant to antibiotics
*foreign DNA interrupted the production of B Galactosidase |
|
|
Term
|
Definition
*single stranded = more unstable
*uracil |
|
|
Term
|
Definition
PCR: need to know exact seq to design primer
DNA RESTRICT: use if need mix frags
*differs b/c need to know seq that needs to be ID (only need to know if seq is present in DNA) |
|
|
Term
|
Definition
* primary struct- AA seq liked polypep bonds
*20 AA
*20^n (n= # residues) |
|
|
Term
|
Definition
*100-1000 AA
*EX: Titin- lrgest known protein (34-350AA)
*diversity is unlimited in theory but in a cell limited to size and composition |
|
|
Term
|
Definition
SIZE
COMPOSITION LIMITATION
POSITION |
|
|
Term
|
Definition
* 40-1000
-in order to fold into right shape for func
-protein synth machinery |
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|
Term
|
Definition
*AA occur @ diff freq
*rare AA become a limiting factor |
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|
Term
|
Definition
*more important than composition
*3D structure is a result of interactions
*Phobic (inside prot), phillic (out, surf) b/c protein in aq soln
EX: Cys
*important b/c 2 Cys residues stabilize protein struct (diS bridge) |
|
|
Term
|
Definition
*has Cys diS bridges between 2 dff chains and w/in same chain
*specific on which Cys wants to bind too |
|
|
Term
|
Definition
*after remove from natural env
CAUSED BY:
1) pH
-need appropriate buffers
2)T
-high T = faster deg
3) presence of protease/nuclease
-protein deg protein
4)absorp to surf
-adhere b/c of air-H2O interface/ glass, plastic
5)storage
-slow oxidation, microbial contamination |
|
|
Term
|
Definition
ENZ COUPLED ASSAY
*Enz-Linked Immunosorbent Assay (ELISA)
SPECTROPHOTOMETRY |
|
|
Term
|
Definition
*2 antibodies
1: specific for target prot and linked on solid support
2: bound to assayable enz; specific to 1st antibod
*[protein] measured indirectly by directly measuring enz activity
*good b/c [protein] use to amplify signal
*quantifies enz activity
*confers ability to measure enz activity & tell how much protein present
*measures enz activity
*can determine how much protein in rxn |
|
|
Term
|
Definition
1) Immobile 1st antibod
2) incubate w/ protein that has sample
2) Add 2nd antibod w/ enz allows to measure
3) Bind, incubation
4) Assay and determine enz activity and convert to initial [protein]
*amount of substrate converted to product indicates amount of assayable protein present |
|
|
Term
|
Definition
*uses Lambert-Beer Law
*calc [protein]
*measures enz activity
*read absorbance on machine and calc [] |
|
|
Term
|
Definition
A: what trying to measure
Io: lihgt intensity
E: extinction coeff
c: []
I: light path in cm (light going through) |
|
|
Term
| Absorbance of Polypeptides |
|
Definition
DO ABSORB IN
*UV (200-400nm)
*b/c lrg extinction coeff in the region of aromatic side chains (Phe, Trp, Tyr)
DON'T ABSORB
*Visible light (400-800nm)
*b/c chromophore in visible light region, [] cant be determined |
|
|
Term
| Protein Purification Characteristics & Strategies |
|
Definition
SOLUBILITY
*salt out
IONIC CHARGE
*Ion Exchange Chromatography
*Electrophoresis
*Isoelectric Focusing
POLARITY
*Phobic Interaction Chromatography
SIZE
*Gel Filtration Chromatography
*SDS-PAGE
BINDING SPECIFICITY
*Affinity Chromatography |
|
|
Term
|
Definition
*purify based on solubility
*salt added to protein soln (ammonium sulfate)
*proteins have diff behaviors and precipitate based on [salt]
*proteins least solub @ pI
SALT IN:
*solub @ low [ion] increases as salt added
*add salt = inc solub
*proteins remain suspended in soln
SALT OUT:
*as [salt] inc = solub dec
*proteins precipitate of of soln |
|
|
Term
|
Definition
*Microbes in milk grow
*if grow too long, acids synth (lactic, acetic)
*cause pH drop and casein precipitation/ coagulation
*casein forms @ pI (pH2) |
|
|
Term
|
Definition
| * interaction of proteins w/ mobile/ stationary phase |
|
|
Term
|
Definition
*total net charge of molecules after separation of ions
*allows to separate molecules easily
*changing buffers pH = manipulation of charge carried by molecule of interest |
|
|
Term
|
Definition
*the pH @ which a molecule has no net charge
pH buff > pI protein
*protein carries -
*+ charged anion exchange resin chosen to capture protein
pH buff < pI protein
*protein carries +
*- cation resin chosen |
|
|
Term
|
Definition
*pH goes above pI = protein has -
-want a anion, +
*protein goes more acidic than pI = cation xΔ
-want a cation, - |
|
|
Term
| Ion Exchange Chromatography |
|
Definition
ANION xΔER
*matrix w/ attached diethylaminoethyl(DEAE)
CATION XΔER
*matrix bearing carboxymethyl(CM) groups
1) mixture w/ 3 proteins
2) stationary matrix has ion xΔer
3) separate based on response to buffer w/ diff [salt] or pH
4) one w/ leat affinity precip out 1st
5) use salt out to dec solub and precipitate out
6) collect eluent in tubes
7) assay [protein]
*separate protein based on attraction ion xΔer but doesn't help find NTerm
*incorporate salt out |
|
|
Term
| Gel (Size Exclusion) Filtration Chromatography |
|
Definition
1) column pacaked w/ large fragments
2) add protein
3) larger protein move faster, sm slower
*small particles have many options to fit through pores, large have less |
|
|
Term
|
Definition
*tells binding specificity characteristics
1) column w/ known matter
2) load w/ something that can bind (ligand , lock and key)
3)protein doesnt bind washed out
4) remove bound protein w/ higher affinity by add compound (must out compete already bound protein)
5) elute
*lock and key analogy |
|
|
Term
|
Definition
*High selectivity, resolution, capacity
*utilizes proteins biological struct or func
*purifications easily achieved |
|
|
Term
Polyacrylamide Gel Electrophoresis
(PAGE)
and
SDS-PAGE |
|
Definition
*helps get pure proteins
*size, mass, weight selective
*sim concept to DNA separation
* - -> + (small move faster)
*proteins move in response to electric field
*small move fast so can determine protein |
|
|
Term
|
Definition
NO SDS: charge of protein maintained
-proteins migrate toward either electrode
-not suitable for molecular weight determination
*W/ SDS & DENATURIZERS: protein w/ -
-protein full denatured and dissociate from eachother
-overall - due to - on SDS
-protein have rod confirmation instead of tertiary shape
*proteins vary in charge so cant move by self or separate so need to add SDS
*sodium dodecyl sulfate- detergent denaturizes proteins; makes linear and adds overall - to protein |
|
|
Term
|
Definition
1) centrifuge mix and see sep based on mass
2) elute
3) centrifuge powerful so allows to separate |
|
|
Term
| Protein Sequencing Techniques |
|
Definition
END GROUP ANALYSIS
- Dansyl Chloride
- Edman Degradation
CLEAVE DIS BONDS
-mercaptan
CLEAVE POLYPEPTIDE CHAINS
-peptidase
-Mass Spectrometry?
REPEATES REMOVAL OF NTERM AA RESIDUE
-Edman Degredation |
|
|
Term
| Protein Sequencing: End Group Analysis |
|
Definition
*proteins composed of subunits need to be spearated
*protein w/ subunits separate and determine every N term AA -> conclude # subunits
*done by: Dansyl chloride forms yellow compound, edman
*determines only 1st AA in NTerm |
|
|
Term
|
Definition
1)Treatment of dansylated polypep w/ aq acids @ high T
2) have DCl w/ OH and polypep
3) form D polypep (gen N term AA bound to DCl: now flourecent)
4) cleavage peptide bond, all other AA celared and free
- 1 flour AA for e. chain on N term
5) separation of dansylated Nterm residue from other AA through chromatography
*do 1x and chain is fragmented
*removes NTerm and peptide bonds
*need time for rxn to occur |
|
|
Term
|
Definition
1) PITC (edmans reagent) reacts w/ NTerm Amino group under mild-alkaline condi
2) PTC adduct treated w/ anhydrous trifluoroacetic acid (cleaves Nterm)
3)extract thializonone AA w/ organic solvent
4) treatment w/ aq acid = more stable PTH AA
*rxn allows to remove only N terminal residues 1@ time
*perform repeated discrimination of AA and NTerm |
|
|
Term
| Protein Sequencing: Cleave DiS Bonds |
|
Definition
*ensures peptides linear
*treat w/ 2 mercaptoethanol (breaks all DiS) / other mercaptan
*sep subunits by break DiS
1) free sulfhydryl groups are alkylated w/ iodoacetate (DiS bond between Cys subunits broke)
2)PREVENTS REFORMATION of DiS |
|
|
Term
| Protein Sequencing: Cleave Polypeptide Chains |
|
Definition
*uses peptidases or chem reagents w/o restriction enz
-peptidase more specific , can select specificity
*endopeptidase -> sm protein frags
*endo/exo peptidase have side chain requirements for scissile pep bond
*specific/non used to recognize bond that want cleaved
*Mass Spec
1) Cyanogen bromide reacts w/ Met
2) cleave peptide bond on CTerm
3) New CTerm residue forms cyclic peptidyl homoserine lactone |
|
|
Term
|
Definition
*bond recognized and cut
Trp: high specificity
-cleaves peptide bonds on CSide of + Arg,Lys if not next to Pro
-allows to cleave specific region in peptide
Pep: nonspecific |
|
|
Term
|
Definition
*get seq based on mass:charge (m/z)
*cleave and run chromatography
1)put protein mix in gas phase
2) vaporize
3) run proteins through vaccuum
4) apply N gas
5)see protein charge depending on Arg, Lys
6)detects mass:charge in proteins
7)differentiate protein based on mass |
|
|
Term
| Protein Sequencing: Repeated Removal of NTerm AA Residue |
|
Definition
| *Edman: only NTerm removed |
|
|
Term
|
Definition
*fermented milk
*digested lactose
*specific bact
*nutrients (Ca, P, K, vit B2,B12) |
|
|
Term
|
Definition
*10000-5000BCE milk incorporated in diet b/c anim
*safe for storing other than drying
-discovered b/c kept in intestinal bags and microbes began to ferment
*medical purposes
*need 3 servings/day
Grigorov: lactic acid bact (tang taste), 20th century
Dannon: 1st yogurt lab/ factory |
|
|
Term
| Yogurt and Lactose Intolerance |
|
Definition
*helps introduce Ca, vit D
*bact already digest lactose so person w/o lactase can ingest w/o problem
*Asian, Af Am, Indian have higher intakes of yogurt because more lactose intolerant |
|
|
Term
|
Definition
1) Enhanced digestability and bioavailability
-beneifts for lactose intolerance
2) addn of probiotics
-benefits gut microflora b/c dec patho
3) FORTIFIED W/ Ca and Vits
-inc Ca intake
-reduced risk of gastrointestinal disease, cardivasc disease, metabolic syndrome, Type2 diabeters, resp disease,
-inc bone, dental health, pregancy |
|
|
Term
|
Definition
Lactobacillus bulgaricus
Streptococcus thermophilus
*sterile env at low T (36C-42C for 48hr) |
|
|
Term
|
Definition
1) milk lactose used as fermentation substrate
2) lactic acid/ other compound form (smell)
3) decrease in pH -> deveopment of undesirable microorg delayed
4) Ca, P in milk converted to soluble form
5)no Ca; majority of proteins digested by proteolytic enz enhances digestibility and overall bioavailability |
|
|
Term
| Phylogenetic Trees and DNA sequencing |
|
Definition
*can be used to tell relatedness
*primary struct from related species resemble one another
*primary struct tells how conserved a seq is, seq can build into tree
1)detect mutations
2)shows relationships of proteins
-Mb and Hb only have 18% relatedness
-but still have similar tertiary struct |
|
|
Term
|
Definition
*sm molecule (153 resid)
*simple ox binding behavior
*8 a-helices connected by short polypep
*heme group interact w/ polypep
*musc rapidly respires under high exertion (high O2)
-diffusion of O2 from caps to tiss is limited by low solub in aq soln (10^4M blood) |
|
|
Term
|
Definition
*reversible binding of O2 so can be used in necessary places
*AA allow reversible bind
PROB: bind other molecules
*CO, NO, H2S can bind to heme groups on protein
*higher affinity than O2 so very toxic
*CO has 200x greater affinity for Hb than O2
EX: Birds in Mine Shaft
*inc in other gases that bind heme |
|
|
Term
|
Definition
*used as O2 storage; [musc Mb] 10x > terrestrial
*needed for diving and long periods under water |
|
|
Term
|
Definition
*bind and release O2
*heterpcyclic ring syst of porphyrin derivate
*in phobic pocket between E & F helix
*4 pyrrole group (A-D) linked by menthene bridge
*has ferrous Fe (FeII) to bind O2
-Fe linked to N on His side chain
*5N total
OXYGENATED Fe(II):
deep purple -> scarlet red (venous vs arterial)
EX: Hb, Mb, CytC |
|
|
Term
|
Definition
1) Pyrrole A
2) Pyrrole B
3) Pyrrole C
4) Pyrrole D
-all have N
5)His F8
-allows reversibly bind
-binds to Fe
6) O2
-forms interaction with His E7 and Fe |
|
|
Term
|
Definition
*Val E11 and Phe CD1 hold heme in place
-allow and restrict access of O2 to protein
-grab and allow His to grab O2
-side chains swing as protein breathes to allow O2 to enter and exit
-change in conformation allows O2 bind and stay bound
*heme between E & F helices w/ phobic interactions
*His E7: interacts w/ O2 by H bond
HISF8 __ Fe -- O2 -- HISE7 |
|
|
Term
|
Definition
BAD
*ferrous Fe of isolated heme is irreversibly oxidized to ferric Fe
-CO molecule bound to heme and color change
-metmyoglobin/methemoglobin formed (brown meat, dried blood)
GOOD
*protein portion of Mb prevents oxidation and allows reversible
*reversible binding: Mb + O2 ⇌ MbO2 |
|
|
Term
|
Definition
Yo2: fractional saturation
-the fraction of O2 bind sites occupied by O2
-0 = no O2 bound to Mb
-1 = all bind sites of Mb taken
pO2: partial press of O2
-ox tension
- [O2] torr
K: dissociation Constant
-value of pO2 at which fractional saturation y=.5
-pO2 @ which 1/2 bind sites occupied
p50: [o2] @ .50 of O2 binding sites occupied
-press at which Mb is 50% sat |
|
|
Term
|
Definition
STANDARD BODY COND Mb:
YO2: .5 @1/2 sat
pO2 = 2.8 torr
*Almost full satu in blood Mb
*arterial: 100 torr, venous: 30 torr
p50= 2.8torr
*simple binding = hyperbolic shape
*Mb like to occupy all bind sites so carry from blood -> musc |
|
|
Term
|
Definition
1) Facilitates O2 diffusion in musc
-major physio role
-efficiently relays O2 from caps to musc cells
-prob b/c low solub so Mb inc O2 solub in musc
2) Oxygen Storage (aquatic mammals)
-Increases O2 solub in musc cells
-diffusion limited by low solub, Mb inc solub |
|
|
Term
|
Definition
*small animals rely on diffusion for O2
*lrg animals (>1mm thick) cant satisfy
-have circ syst w/ O2 binding molecule (Hb, hemerythrin, hemocyanin) to boost limited solub of O2 in H2O
-need syst to inc solub in H2O |
|
|
Term
|
Definition
*don't have heme group, only in invertebrate
-'heme' referring to in the blood
HEMERYTHRIN
colorless -> violet,pink b/c O2 states of bind molecule (when oxygenated)
HEMOCYANIN
colorless -> blue when binds to O2
extracellular transport |
|
|
Term
|
Definition
*delivers O2 to various tiss
*O2 bind explains control of enz act
*tertramer (a2B2) consisting of 2 dimers of aB protomers |
|
|
Term
|
Definition
1) O2 bind
2) conformational change (protomer rotate 15*)
3) change cause interaction of subunits and AA
-most interactions phobic allows for proper bind
4) shatters deoxyhemoglobin crystals when exposed to Ox
5) B subunits closer together
*associations have extensive contacts, mostly phobic |
|
|
Term
| O2 Binding Behavior of Hb |
|
Definition
*sigmoidal shape
-coop bind, blood delivering/ releasing more O2 to tiss than Mb
*less affinity for O2 than Mb
-Mb is able to pull away O2 molecules from heme (high association w/ O2 (tight)
*increase in slope
-caused by affinity increase after 1st O2 bind |
|
|
Term
|
Definition
STANDARD BODY COND Hb:
YO2: .95 @ full sat of arterial O2 (all bind sites occupied)
pO2 = 100 torr
YO2: .55 @ 1/2 sat of venous O2
pO2: 25 torr
p50= 26 torr
*cooperative binding = sigmoidal shape |
|
|
Term
|
Definition
*one encourages bind of another
*as more O2 binds, affinity for next increases
*binding of 1 ligand affects binding of additional ligands to another
*O2 bind to 1 subunit increases O2 binding of remaining
-Hb subunits compete for first O2 |
|
|
Term
|
Definition
*describes the degree of sat of Hb (func of pO2)
*describes O2 binding curve of Hb |
|
|
Term
|
Definition
n=1
*noncooperative O2 binding
*bind of O2 is independent (won't have an affect on the other)
n>1
*positively cooperative
*bonding of 1 O2 increases affinity of Hb for further binding
n<1
*negatively cooperative
*O2 binding reduces affinity of Hb for bind other O2 |
|
|
Term
|
Definition
T
*deoxyHb
*unoccupied, no O2 bound
*4 hemes ABOVE heme plane (near His F8)
R
*oxyHb
*Fe(II) moves into center plane
*T->R conformational change when O2 bind
-both states stable
*H bond between AA change depending on T or R |
|
|
Term
|
Definition
T -> R
*pK dec (N term, C term of a/B residues)
*more acidic, more likely to give up protons
*pH drop, inc [H] (shifts affinity for O2)
*Hb release .6 proton for each binding O2 |
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Term
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Definition
1) Inc pH cause inc in affinity for Hb
-affinity (of O2 for Hb) inc as pH inc ([H] dec)
-vice versa
-allows Hb to release & bind O2 in lung, musc, resp tiss
2) Produced CO2 diffuses slowly from resp tiss to caps; dissolved CO2 slowly forms BiCarb
-important role in o2/CO2 exchange between lung and resp tiss
- LUNG HbO2 affinity < MUSC MbO2 affinity
*provides the mech to provide additional O2 in highly active musc (pO2 <20torr) |
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Term
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Definition
*in erythrocytes
*accelerates formation of biCarb
*need act b/c w/o -> inc [Co2] have bubble is blood
-instead seen as BiCarb b/c of H2O |
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Term
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Definition
pO2 low
*HCO3- formation gen heat
* Hb in T state takes H
*form T stateinduces Hb to release O2
*H+ uptake facilitates CO2 transport by stim HCO3 form
*equil: HbH removes proton from HCO3- +H and allows to push towards lungs |
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Term
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Definition
pO2 HIGH
*T->R triggers release of H (ion pair) so now Hb can take up O2 |
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Term
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Definition
*normally, no change in blood pH caused by Hb->Hbx exchange
-lactic acid causes pH decrease
*highly act musc generate lac acid
-lac acid -> pH drop -> more O2 released (sm pH = more P2 release) |
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Term
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Definition
*form w/ CO2 so CO2 does not exist as CO2 in blood
*forms at Nterm AA
*THb bind more carbamate CO2 than R
*protons released at formation
-protons promote O2 release (Bohr Effect)
high [Co2] -> favor T state -> release O2 |
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Term
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Definition
*protein that affecs the affinity of Hb (decreases affinity)
-pure Hb higher affinity > blood Hb b/c CO2 and other components affect
-allows transfer of maternal, fetal blood
MAMMALS
*decrease Hb affinity by keeping it in deoxy form
VERTS
*other compounds that give same affect |
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Term
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Definition
*binds to deoxyHb and keep in T state to dec O2 affinity
*binds tightly to deoxyHb, weakly to oxyHb
ART: pO2= 100, Hb sat w/ O2= 95%
VEN: pO2= 30, Hb sat w/ O2= 55%
*in caps, Hb unloads 40% bound O2 |
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Term
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Definition
*Hb affinity inc -> little bound O2 released
*curve at lower pO2 |
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Term
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Definition
*helps supply fetus w/ O2 (from maternal circ by placenta)
*[BPG] same in adult, fetal
-adult BPG binds tighter to Hb
-fetal has high BPG affinity, maternal has low
-low BPG affinity (maternal)= high O2 affinity
*b/c fetal has higher O2 affinity, facilitated transfer of O2 to fetus |
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Term
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Definition
*subunit composition of a2y2 (adult= a2B2)
*y changes to B
*His residue in B chain form interact w/ BPG; His stabilizes BP-deoxyHb complex
*y has uncharged Ser which allows low affinity in maternal |
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Term
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Definition
*colorless/odor/taste, non irritating gas
*leading cause death
*can kill fast or slow (35ppm for 8hr w/o effects) |
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Term
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Definition
*inhaling combustible fumes
-generators, lanterns, gas ranges, etc
*inhaling smoke from closed place fire |
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Term
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Definition
*causes hypoxia at tiss level
*CO binds to heme -> O2 cant bind (200x > affinity) -> form carboxyHb (COHb) -> less O2 carried
*when binds to cardiac Mb, less O2 available for aerobic metab -> dec cardiac contract, less O2 transport (AKA low flow hypoxia) |
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Term
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Definition
*every body syst (heart, brain, lung sens)
*causes headache, nausea, vomit, fatigue
*cause low flow hypoxia
*poisoning worse when w/ other cond
*blood, O2 content redby 1/2 w/ COHb |
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Term
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Definition
*ID symptoms
*COHb levels:
-danger if > 25% COHb
-lethal if > 50% COHb
*measure ven,art blood gases
-pO2 levels
*complete blood count assess
*lactate levels
*cardiac enz levels
*admin O2 @ 15L/min (4hr)
-red 1/2 life COHb (300-90min)
*severe cases:
-intubate -> ICU, monitor heart -> cardiovasc resp support -> meurologic development -> HBO therapy (heart chamber oxygenation) = red 1/2 life COHb (300-30min) |
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Term
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Definition
*CO detector
*professionally installed fuel burning appliance
*vented space heaters |
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