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Myoglobin
• Globular protein, 3° folding, 8 right handed α helices (A – H) with 7-20 AAs each
• Heme – prosthetic group @ hydrophobic crevice; porphyrin ring + central Fe
• Nomeclature – letter of helix + AA position; order of AA residue
• 5 non-helical segments – AB, CD, EF, FG, GH
• 2 non-helical terminal segments – NA1 and NA2, HC1-HC5
Hemoglobin
• Quaternary structure
• Tetramer = 2α and 2β subunits facing each other across a central cavity
• Packing contact @ α1β1 & α2β2, fixed
• Sliding contact @ α1β2 & α2β1, not fixed
• 4 heme groups = 4 oxygen binding sites
• O2 binding @ 6th coordination position of heme
• Val E11 pushes and destabilizes nonperpendicular molecules @ binding plane (i.e. CO) allowing reversible binding
• Conserved AA:
o His F8 (proximal His / residue 93) - always bound to Fe
o His E7 (distal His / residue 64) - binds to O2, then O2 binds to Fe
o PheCD1, Leu F4 – heme contact
o Gly B6 – allows close approach of helices B and E via B6&E8
Heme
• Prosthetic group in hydrophobic pocket for heme sequestration and prevention of Fe2+ oxidation to Fe3+
• Protoporphyrin ring – organic, 4 pyrroles via methenyl bridges
• Fe2+ atom - @ center or porphyrin ring
• 6 ligands via coordinate bonds
o Four are 4N of porphyrin ring
o 5th – HisF8 or residue 98 or proximal His
o 6th – reserved for oxygen during oxygenation
Hemoglobin Conformations
• T / taut = lower O2 affinity = deoxyhb = dome shaped
• R / relaxed = higher O2 affinity = oxyhb
• Transition from T to R
o 15° rotation of α2β2 relative to α1β1
o Breaking of 8 salt bridges that stabilize T state
o New H bonds @ α1β2 interface = AspG1-AsnG
o Molecular rearrangments – O2 pulls heme Fe straightening of dome
• molecular strain @ HisF8 from 8° to 0°, Val F65 pushed away
Carbon Monoxide
• Forms carboxyhemoglobin, by product of heme degradation, 1% of O2 binding sites
• Heme affinity = 25000x than O2
• Can interfere with O2 acquisition by heme
• Shifts OBC left
• HisE7 reduces affinity of linear CO binding to heme
• Optimal binding orientation= 121°
Methemoglobinemia
• Hemoglobin Fe2+ oxidized to Fe3+
• Hb cannot carry O2
• Results when metHb > 1.5%
• Leftward shift
Hemoglobinopathies
• Inherited, substitution or absence of AA in globin chain
• Sickle Cell Anemia – point mutation @ β chain, Glu6 Val = HbS
o Val6 on β chain produces sticky patch at exterior
o Complimentary site @ deoxy HbA or HbS + sticky patch = aggregation = RBC distortion, HbS stabilization
o ↓ pO2 = deoxyHbS aggregation = sickling
o Clinical symptoms: hemolytic anemia, infarctive crisis via occlusion, P50=31 (rightward shift = ↑ 2,3 DPG)
o Oxygenation = ↑ oxyHbS = ↓ deoxyHbS = mask complimentary site = ↓ sickling
o Deoxy HbA from transfusion = no sticky patch
Thalassemia Hb Info
defective gene expression of α globin
α-Thalassemia 2 Silent carrier, one gene deletion
α α-Thalassemia Trait Hb Barts (γ4) 2 gene deletion
Hemoglobin H Hb H (β4) 3 gene deletion
Hydrops Fetalis No α chains
Minor β/β+ to β/β° Mild hypochromic microcytic anemia, asymptomatic
β Intermedia β°/β+ to β+/β+ depends on β+ severity
Major β°/β+ to β°/β° severe anemia, multiple organ damage, transfusion-dependent
Transport
• Same as O2 transport:
• Convection transport = tissue lungs
• Diffusion from ↑ pCO2 (venous blood) ↓ pCO2 (lungs)
• Differential solubility of O2 and CO2
• Forms in Blood:
Form Amount Info
Dissolved CO2 9-10% (3°) CO2 = 20x more soluble than O2
75-78% (1°) Carbonic anhydrase reaction: CO2 enters RBC + H2O + carbonic
HCO3-
anhydrase = H2CO3 spontaneously dissociates into HCO3- + H+
Carbaminohemoglobin 13-20% (2°) CO2 + amino group of hemoglobin
Carbaminohemoglobin Formation
• CO2 + terminal NH2 of aliphatic CHONs
• RBC pH = 7.2 = NH2 NH3
• Removal of some uncharged NH2 via carbamino formation generate more NH2 + H+
• Hemoglobin is most important:
o More Hb than CHON
o Tetramer = 4 terminal NH2
o DeoxyHb (venous blood) forms carbamino more readily than oxyHb oxygenation = CO2 release
Regulation
• Buffering – HCO3- and H2CO3 buffering in ionizable groups (i.e. 4 N-terminal NH2 groups, imidazole of His38, phosphate groups)
• Isohydric Mechanism – Hgb can take up H+ from CO2 without pH change
Haldane Effect
• H+ affinity: deoxyHb > oxyHb
• O2 + Hb = ↓ CO2 affinity (CO2 dissociation curve rightward shift, Haldane effect)
• CO2 + Hb = ↓ O2 affinity (Bohr effect)
Chloride Shift
• Exchange of HCO3 and CL between plasma and RBC = maintain electrical neutrality
• cross membrane via Band 3