绿色荧光蛋白（GFP）的CPL光谱测量

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圆偏振发光（CPL）测量了手性化合物的左、右旋圆偏振光发射强度的差异。与CD不同，CPL提供了关于手性分子激发态性质的信息。绿色荧光蛋白（GFP）是一种在暴露于紫外线时发出绿色荧光的蛋白质，并且已被证明具有手性性质。Application Note CPL Spectrum Measurement ofGreen Fluorescent Protein (GFP)2/2Application Note CD-0030 CPL Spectrum Measurement of Green Fluorescen t Protein (GFP ) Introduc t ion Circula r ly polarized l uminescence (CPL) measures the difference in the em i ssion i n tensit i es of left- a n d r ight-ha n ded circularly polarized l i ght of chiral compounds. Unlike CD， CPL provides informa t ion on t h e excited state properties of chiral molecules . CPL-300 Ci r cularly Po l a r ized Lumi n escence Spect r o p hotomete r Green f luoresce n ce p rotein (GFP) is a protein that emits g r een fluorescence when exposed to ultraviolet light and has been shown to h ave chiroptical propert i es. T h is application note illustrates the CPL and CD spectra of GFP obtained with the CPL-300 and J-1500， respectively. Keyword s J-1500， Ci r cu l ar d i c h roism ， C PL -300， Circ u la r l y p o l ar i ze d luminescence， Pro t ein str u ctu r e ， Flu oresce n ce ， B i ochemist r y Exper i m e ntal Measurement Conditions CPL-300 Circularly Polarized Luminescence Excitation Wavelength 399nm Excitation Bandwidth 12nm Data Interval 1nm Response 16 seconds Emission Bandwidth 8nm Scan Speed 10 nm/min Accumulations 36 Path Length 10mm Measurement Conditions J-1500 Circular Dichroism Spectrophotometer Data Pitch 0.2 nm D.I.T. 1 sec Bandwidth 1nm Scan Speed 200 nm/min Accumulations 16 Path Length 20 mm R esults CD and absor p tio n spect r a (Figu r e 1， g r een) of a 0.03 mg/mL GFP solu t i o n was measured in the nea r -UV and visible r e gion s . T h e GFP ch r omophor e is obs er ved in t h e visi b le r e gion a t 400 nm and th e absor p t i on b a n ds d u e t o t he aromatic ami n o acid resid u es are o b s e rv e d in th e n ear -UV r e gion betwe e n 250 and 300 nm. The CPL and fluor e scence s pe ctra were also obta in ed and show in b lue i n Figu r e 1. The c h aracteristic fl u o rescen t and CPL s p ectra of GFP a r e o b s er ved b e twe e n 470-570n m. Figure 1. C P L (b l u e) a n d C D (g ree n) s p e c tr a (top) o f g r ee n f l u o re sc e n t p r ot e i n . Th e a b so r p t ion (gr ee n ) a n d f l u or e sc en c e (bl u e)spectra ar e shown at the bot t om . R efer e nces 1. Hiromasa Goto， Isao Sawada， and Nobuhiko Nomura， Internat i onal Jour n al of Polymeric Mater i als， 2010， 59， 786-792 JASCO INC. 28600 Mary's Cour t ， E a s to n ， M D 21601US A