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20231223065649.0 |
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231223s2004 xx ||||| 00| ||eng c |
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|a pubmed24n0514.xml
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|a (DE-627)NLM154266264
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|a (NLM)15773127
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|a DE-627
|b ger
|c DE-627
|e rakwb
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| 041 |
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|a eng
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| 100 |
1 |
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|a Ichikawa, Sosaku
|e verfasserin
|4 aut
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| 245 |
1 |
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|a Phospholipase D-mediated aggregation, fusion, and precipitation of phospholipid vesicles
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| 264 |
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|c 2004
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|a Text
|b txt
|2 rdacontent
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|a ohne Hilfsmittel zu benutzen
|b n
|2 rdamedia
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| 338 |
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|a Band
|b nc
|2 rdacarrier
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|a Date Completed 03.02.2006
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|a Date Revised 26.10.2019
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|a published: Print
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|a Citation Status MEDLINE
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| 520 |
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|a Large unilamellar vesicles with a diameter of 100 nm were prepared from the zwitterionic phospholipid POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) at pH 8.0. After addition to these vesicles of the enzyme phospholipase D (PLD) from Streptomyces sp. AA586 at 40 degrees C, the terminal phosphate ester bond of POPC was hydrolyzed, yielding the negatively charged POPA (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidic acid) and the positively charged choline. While the reaction yield in the presence of 1 mM Ca2+ reached 100%, the yield was only approximately 68% in the absence of Ca2+. Furthermore, in the absence of Ca2+, the size of the vesicles did not change significantly with time upon PLD addition, as judged from turbidity, dynamic light scattering, and electron microscopy measurements. In the presence of 1 mM Ca2+, however, PLD addition resulted in vesicle aggregation, fusion, and precipitation, originating from the interaction of Ca2+ ions with the negatively charged phospholipids formed in the membranes. Vesicle fusion was monitored by using a novel fusion assay system involving vesicles containing entrapped trypsin and vesicles containing entrapped chymotrypsinogen A. After vesicle fusion, chymotrypsinogen A transformed into a-chymotrypsin, catalyzed by trypsin inside the fused vesicles. The alpha-chymotrypsin formed could be detected with benzoyl-L-Tyr-p-nitroanilide as a membrane permeable chymotrypsin substrate. The observed vesicle precipitation occurring after vesicle fusion in the presence of 1 mM Ca2+ was correlated with an increase of the main phase transition temperature, Tm, of POPA to values above 40 degrees C
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|a Journal Article
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|a Research Support, Non-U.S. Gov't
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|a 1-palmitoyl-2-oleoyl-glycero-3-phosphatidic acid
|2 NLM
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|a Cations, Divalent
|2 NLM
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|a Lipid Bilayers
|2 NLM
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|a Phosphatidic Acids
|2 NLM
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|a Phosphatidylcholines
|2 NLM
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|a Phospholipids
|2 NLM
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|a Chymotrypsinogen
|2 NLM
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| 650 |
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|a 9035-75-0
|2 NLM
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| 650 |
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|a Phospholipase D
|2 NLM
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| 650 |
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|a EC 3.1.4.4
|2 NLM
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|a Trypsin
|2 NLM
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|a EC 3.4.21.4
|2 NLM
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|a Calcium
|2 NLM
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| 650 |
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|a SY7Q814VUP
|2 NLM
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| 650 |
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|a 1-palmitoyl-2-oleoylphosphatidylcholine
|2 NLM
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| 650 |
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|a TE895536Y5
|2 NLM
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| 700 |
1 |
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|a Walde, Peter
|e verfasserin
|4 aut
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| 773 |
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|i Enthalten in
|t Langmuir : the ACS journal of surfaces and colloids
|d 1992
|g 20(2004), 3 vom: 03. Feb., Seite 941-9
|w (DE-627)NLM098181009
|x 1520-5827
|7 nnns
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| 773 |
1 |
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|g volume:20
|g year:2004
|g number:3
|g day:03
|g month:02
|g pages:941-9
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|a GBV_USEFLAG_A
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|a SYSFLAG_A
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|a GBV_NLM
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|a GBV_ILN_22
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|a GBV_ILN_350
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|a GBV_ILN_721
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|a AR
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|d 20
|j 2004
|e 3
|b 03
|c 02
|h 941-9
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