Photo-dynamics and thermal behavior of the BLUF domain containing adenylate cyclase NgPAC2 from the amoeboflagellate Naegleria gruberi NEG-M strain

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• 作者：A. Penzkofer^a ; ^{alfons.penzkofer@physik.uni-regensburg.de} ; M. Tanwar^b ; S.K. Veetil^b ; S. Kateriya^b ; M. Stierl^c ; P. Hegemann^c
• 关键词：Photo-activated cyclase ; BLUF domain ; Amoeboflagelate Naegleria gruberi ; Flavin ; Photo-cycle dynamics ; Photo-degradation ; Concentration dependent protein color center formation ; Apparent protein melting temperature ; Temperature dependent protein melting t
• 刊名：Chemical Physics
• 出版年：2013
• 出版时间：1 February, 2013
• 年：2013
• 卷：412
• 期：Complete
• 页码：96-108
• 全文大小：1301 K

文摘

The absorption and emission spectroscopic behavior of the photo-activated adenylate cyclase NgPAC2 from the amoeboflagellate Naegleria gruberi NEG-M strain was studied in the dark, during blue-light exposure and after blue-light exposure. The typical BLUF domain (BLUF = Blue Light sensor Using Flavin) flavin cofactor absorption and fluorescence photo-cycle dynamics was observed. For fresh samples a reversible concentration dependent protein oligomerization occurred showing up in free flavin binding and protein color center formation with increasing protein concentration. Thermal and temporal irreversible protein unfolding with loss of BLUF domain activity was investigated. Temperature dependent protein melting times and the apparent protein melting temperature were determined. The photodynamic behavior of the NgPAC2 is compared with the behavior of the previously investigated photo-activated cyclase NgPAC1 (nPAC) from the same N. gruberi NEG-M strain.