Plastocyanin microheterogeneity in Scenedesmus acutus MT8
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Title |
Plastocyanin microheterogeneity in Scenedesmus acutus MT8
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Creator |
Dimitrov, Mitko I
Donchev, Anthony A Toromanov, Georgi R Getov, Vladimir I Toncheva-Panova, Tonka G |
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Subject |
Plastocyanin
Microheterogeneity Dimorphism Isoelectric focusing Chromatofocusing |
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Description |
310-316
Two total plastocyanin (PC) fractions — loosely bound (lPC) and strongly bound (sPC) were extracted (84% and 16%, respectively) from the homogenate of Scenedesmus acutus MT8. Two-fold isolation-purification procedure including DE-52 chromatography separated lPC into a smaller oxidized [lPC (II)] and a larger reduced [lPC(I)] fractions, in contrast to sPC, where sPC(II) greatly dominated over sPC(I). Analytical isoelectric focusing (IEF) separated lPC(II) into two main fractions only in the presence of 8 M urea, implying microheterogeneity. Preparative IEF in immobiline pH-gradient of 3.2-4.1 separated lPC(II) into two blue fractions – a more alkaline lPC'(II) and a more acidic lPC''(II), which were probably stereoisomers. Their UV-Vis spectra exhibited rarely observed tryptophane (291.5 nm) and some differences at 270 and 287 nm. The exact molecular masses of apo-/holo-lPC (10131 Da/10194 Da) were determined by mass spectrometry. The number of -SH groups was determined from the mass difference between alkylated with 4-vinylpyridine (4-VP) and nonalkylated protein. Additionally, a simple procedure for simultaneous separation of both primary structure and stereoisomers of PC was developed. |
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Date |
2008-11-06T04:09:44Z
2008-11-06T04:09:44Z 2008-10 |
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Type |
Article
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Identifier |
0301-1208
http://hdl.handle.net/123456789/2372 |
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Language |
en_US
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Publisher |
CSIR
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Source |
IJBB Vol.45(5) [October 2008]
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