The Main (Glyco) Phospholipid (MPL) of Thermoplasma acidophilum
Abstract
:1. Introduction
2. Growth of Thermoplasma
3. Cell Lysis Experiments
4. Composition of Membrane Lipids
5. Differential Thermal Analysis (DTA) and Differential Scanning Calorimetry (DSC)
5.1. Cryoprotectants
5.2. Mixture of MPL with Other Lipids
5.3. Mixture with Cholesterol
5.4. Mixture with Pore-forming Peptides and Ionophores
6. Model Membranes
6.1. Black Lipid Membranes
6.2. Monomolecular Films at Water/Air Interface
6.3. MPL Liposomes
6.4. Mixed Liposomes
6.5. Proton Permeability and Light-Driven Proton Pumping
6.5.1. Proton Permeability
6.5.2. Reconstitution of Bacteriorhodopsin (BR) and Light-Driven Proton Pumping
6.5.3. Light-Driven Liposomal ATP Synthesis
7. Applications
Funding
Patents
Conflicts of Interest
Abbreviations
AFM | atomic force microscope |
ATP | adenosine-triphosphate |
BLM | black lipid membrane |
BR | bacteriorhodopsin |
CF | carboxyfluorescein |
DHMG | 1,2-dihexadecyl-3-O-ß-D-maltosyl-sn-glycerol |
DMPC | dimyristoylphosphatidylcholine |
DMSO | dimethyl-sulfoxide |
DOC | deoxycholate |
DOPC | dioleoylphosphatidylcholine |
DPGG | dipalmitylglucosylglycerol |
DPhG | diphytanylglycerol |
DPhGG | diphytanylglucosylglycerol |
DPhPC | diphytanylphosphatidylcholine |
DPPC | dipalmitoylphosphatidylcholine |
DPPG | dipalmitoylphosphatidylglycerol |
DSPC | distearoylphosphatidycholine |
DSM | Deutsche Sammlung von Mikroorganismen |
DSC | differential scanning calorimetry |
DTA | differential thermoanalysis |
EL | egg (yolk) lecithin |
EM | electron microscopy |
EPR/ESR | electron paramagnetic resonance/electron spin resonance |
FCCP | carbonylcyanide-4-(trifluoromethoxy)phenylhydrazone |
GDNT | glycerol-dialkyl-nonitol-tetraether |
GL | glycolipid |
LPS | lipopolysaccharide |
MGL | main glycolipid |
ML | Micrococcus luteus lipid extract |
MPL | main glycophospholipid from Thermoplasma acidophilum |
OD | optical density |
Palm | palmitic acid |
PBFI | potassium-binding (benzofuran) fluorescence indicator |
RT | room temperature |
SBL | soybean lecithin |
SDS | sodium dodecyl-sulfate |
TDOC | taurodeoxycholate |
TEL | tetraether lipid |
T.a. | Thermoplasma acidophilum |
TPLF | total polar lipid fraction |
TPLF | Ta total polar lipid fraction from T.a. |
Val | valinomycin |
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pH | 1.2 | 1.5 | 1.8 | 2.4 | 3.0 |
---|---|---|---|---|---|
Growth Temperature | |||||
39 °C | 2.8 [22,24] | ||||
40 °C | 3.2* [20] | ||||
45 °C | 3.6 [21] | ||||
50 °C | 3.8* [20]; 3.9 [21] | ||||
55 °C | 4.0 [21] | 2.8 [18]; 4.1 [21] | 4.8 [21] | 5.1 [21] | |
59 °C | 3.8 [22,24] | ||||
60 °C | 4.2* [20]; 4.5 [21] |
Growth at 59 °C; Lipid Fraction From T.a. | Glass Transition Range [°C] | Metastable Phase Transition Range [°C] | Main Phase Transition Range [°C] | Tm [°C] | ΔH [J/g] |
---|---|---|---|---|---|
Total lipid | −95/−50 | −40/−10 | −16.5 | 5.0 | |
Phospholipid | −95/−50 | −34/−8 | −14.3 | 8.9 | |
MPL | −95/−50 | −32/−7 | −14.0 | 8.9 | |
Glycolipid | −95/−65 | −65/−30 | −30/−13 | −17.7 | 9.0 |
MPL | 39 °C | 49 °C | 59 °C | ||||||
---|---|---|---|---|---|---|---|---|---|
Scan | dT/dt [K/s] phase | °C | endo-/exo-therm | dT/dt [K/s); phase | °C | endo-/exo-therm | dT/dt [K/s]; phase | °C | endo-/exo- therm |
heating | 0.02–0.13 | −30 → +30 | endo-exo-endo | 0.017–0.083 | −30 → +20 | endo-exo-endo | |||
Tm | A→C | −15 | endo | A→C | −15 | endo | A→C | −15 | endo |
A/C→B’ | 0 | exo | A/C→B’ | ~ 0 | small exo | ||||
B’→C | 20 | endo | B’→C | 20 | small endo | +17 | endo | ||
ΔH | depends on dT/dt | depends on dT/dt | widely indep-endent | ||||||
cooling | 0.08 C→A | −20 → −45 | exo | 0.033 C→A | −30 → −50 | exo | 0.033 C→A | exo | |
Tm | −27 | exo |
Phase Transition | From | to | Mixture | ||||
---|---|---|---|---|---|---|---|
Tm [°C] range | J/mol | kJ/mol K | |||||
MPL | Glass transition | −90/−50 | 1.26 | Rigid glass | Gel analogue | ||
MPL | Main transition | −30 | −14 | Gel analogue | Liquid crystalline | ||
DPhGG | Glass transition | −84/−61 | 0.74 | Rigid glass | Liquid crystalline | Unlimited, no phase separation | |
DPGG | sharp | 62 | −98 | Limited; only >62 °C; <62 °C phase separation with DPGG-rich and DPGG-poor domains | |||
DPPG | Transition and mixture behavior similar to DPGG | ||||||
DPPC | weak pretr, sharp ph.tr. | 35/ 42 | Limited, phase separation with DPPC-rich (ph.tr. >0 °C) and DPPC-poor (ph.tr. at −13 °C) domains |
Shift of Tm [°K] | MPL/Ionophore Ratio at ΔH = 0 | Times 2# | DPPC/Ionophore Ratio at ΔH = 0 | ||
---|---|---|---|---|---|
MPL | DPPC/DHMG* | ||||
Alamethicin | −4.7 | −4.2 | 0.5 | 1.0 | 1.5 |
Melittin | −15.8 | −10 to −20 | 2–2.5 | 4–5 | 6–8 |
Valinomycin | >0 | −0.6 | 0.5 | 1.0 | 0 |
Nonactin | −3.4 | −6.7/−2.7* | ~0.3 | ~0.6 | ΔH increases |
Lipid | Solvent | Cm [µF/cm2] | td [nm] | Stability | Conductance [µS/cm2] | ||
---|---|---|---|---|---|---|---|
Valinomycin 10−7 M | Nonactin 10−6 M | Gramicidin 5 × 10−11 M | |||||
MPL | n-decane or squalene | 0.744 | 2.5 (3.0) | + | 200–250 | 300 | 60 |
MGL | 2.5 (3.0) | +/(+)* | 700 (1000)* | 110 | |||
GDNT | ~0.7 | 2.5–3.0 | + | 1200 | n.d. | 130 | |
DPhGG | n-decane | 0.412 | ~4.5 | (+) | 125 | ||
squalene | ~0.7 | 2.6–3.0 | − | 900 | |||
DPhPC | n-decane | 0.412 | 4.72 5.68 | + | 185 85 | 50 | 40/double conc. 100 |
squalene | ~0.7 | ~3.0 | (+) | 480 | 1100 |
Preparation | Mean Diameter [nm] ± SD | Dispersity Index 0-1 | Reference [62] |
---|---|---|---|
Hand-shaken | 2500 to > 7500 | 0.9 | [63] |
Sonication | 600 ± 40 | 0.5 | [64] |
Detergent dialysis | 370 ± 35 | 0.5 | [65] |
French pressure cell | 151 ± 22 | 0.4 | [66] |
Polycarbonate filter extrusion 200 nm | 221 ± 63 | 0.6 | [67] |
Polycarbonate filter extrusion 100 nm | 120 ± 40 |
Spin Probe | Lecithin Liposomes | MPL Liposomes |
---|---|---|
3-Doxyl-5α-cholestane | 0.655 | 0.932 |
5-Doxylstearic acid | 0.644 | 0.781 |
16-Doxylstearic acid | 0.201 | 0.258 |
5-Doxyldecane | 0.130 | 0.187 |
di-tert-Butylnitroxide | 0.117 | 0.091 |
Liposomes | Egg Lecithin | MPL | ||||
---|---|---|---|---|---|---|
Original | Bile Acid Salts | Original | Bile Acid Salts | |||
10 mM | 30 mM | 10 mM | 30 mM | |||
size | 159.8 nm | 47.2 nm | 1898.4 nm | 131.5 nm | 107.9 nm | 114.8 nm |
Lipid/Detergent Ratio | MPL | MPL/EL 75:25 (% g/g) | MPL/EL 50:50 (% g/g) | MPL/EL 25:75 (% g/g) | Egg Lecithin |
---|---|---|---|---|---|
0.05 | 372 ± 98 | 558 | − | 490 | − |
0.1 | 446 (396–495) | 396 (199–550) | 728 (359–778) | 484 (422–645) | − |
0.15 | n.d. | 263 (169–307) | 431 (386–664) | 471 (453–489) | − |
0.2 | 444 (421–466) | 461 (480–792) | 818 | 413 (325–510) | 153 ± 5 (145–162) |
0.3 | − | − | − | − | 144 ± 9 (137–150) |
Days | MPL | MPL/EL 75:25 (%g/g) | MPL/EL 50:50 (%g/g) | MPL/EL 25:75 (%g/g) | Egg Lecithin | |
---|---|---|---|---|---|---|
0–2 | 466 | 372 | 408 | 386 | 159 | |
5–10 | 450 | 394 | 396 | 146 | ||
15–38 | 358 | 695 (day 38) | 395 | 411 | 149 | |
49–70 | 446 | − | 366 (day 49) | 405 | 149 (day 66) | |
80–90 | − | − | 446 (day 89) | − | ||
128 | 434 | − | − | - | − | |
746 | 498 | − | − | - | − |
Lipid | 4 °C | 30 °C | 50 °C |
---|---|---|---|
Egg lecithin | 9% | 41% | 100% |
DPPC | 6% | 32% | 100% |
DPPC/MPL molar ratio 2:1 | 5% | 8% | 62% |
DPPC/MPL molar ratio 1:2 | 3% | 5% | 15% |
MPL | 2.5% | 3% | 8% |
Liposomes | MPL | Egg Lecithin | Remarks |
---|---|---|---|
+ 5µM Val | + 5µM Val | ||
Valinomycin | 4.5 × 10−6 × cm sec−1 | 10−5-10−4 × cm sec−1 | Proton permeability coefficient |
+ Val | − | + | excess valinomycin |
+ Palm | − | ++ | |
+ FCCP | +++ | +++ | |
2.7 × 10−5 nmol H+ sec−1 cm−2 | 1.2 × 10−4 nmol H+ sec−1 cm−2 | Proton permeability |
Addition | Concentration | Initial Pump Rate | Proton Efflux [ng H+ x min−1] | |
---|---|---|---|---|
Calculated per mg MPL | Calculated per mg BR | |||
nil | 1.4 | 0.6 | 0.1 | |
valinomycin | 0.24 µM | 41.6 | 1.2 | 0.15 |
Val + gramicidin | 0.24 µM + 1 µM | 15.8 | 0.05 | 0.008 |
Val + gramicidin | 0.24 µM + 5 µM | 6.2 | 0.02 | 0.003 |
Val + FCCP | 0.24 µM + 1 µM | 15.5 | 0.21 | 0.035 |
Val + FCCP | 0.24 µM + 6 µM | 11.6 | 0.58 | 0.1 |
Val + FCCP | 0.24 µM + 15 µM | 6.3 | 0.58 | 0.1 |
Lipid | Liposomes Size [nm] | nmol ATP (mg × min)−1 | Detergent/Method | Reference |
---|---|---|---|---|
MPL | 132 | 13.4 | Octylglucoside det-dialys/ French press | [65,66,72] |
SBL | 52 | 77.1 | Taurodeoxycholate (TDOC) | [77,78,79,80] |
DOPC | 86 | 40.5 | ||
ML | 123 | 2.2 | ||
DMPC | 120 | 0 |
© 2019 by the author. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
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Freisleben, H.-J. The Main (Glyco) Phospholipid (MPL) of Thermoplasma acidophilum. Int. J. Mol. Sci. 2019, 20, 5217. https://doi.org/10.3390/ijms20205217
Freisleben H-J. The Main (Glyco) Phospholipid (MPL) of Thermoplasma acidophilum. International Journal of Molecular Sciences. 2019; 20(20):5217. https://doi.org/10.3390/ijms20205217
Chicago/Turabian StyleFreisleben, Hans-Joachim. 2019. "The Main (Glyco) Phospholipid (MPL) of Thermoplasma acidophilum" International Journal of Molecular Sciences 20, no. 20: 5217. https://doi.org/10.3390/ijms20205217