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trytophan fluorescence spectroscopy for protein structure
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trytophan fluorescence spectroscopy for protein structure# Biology - 生物学
X*7
1
三年后,十一岁的新主人第一次让我尝到了鲜血。
“怕什么?杀人又怎么样呢?那些人和猪狗有什么区别?……反正我没有亲人,反
正没人说我做的对不对,反正我只是没人要的孩子。”十岁的主人看着尸体冷冷地笑,
我听见了她内心这样的话。
“任何人都不会在乎我,那么我也不会在乎任何人……”
“我绝对不会为任何人哭。”
在杀人时,我不停地听见她内心这样地反复着。
杀戮之门一开,走进去就永无回头之路,一直到死。
命运……如果真的有人类所谓命运的话,那么命运的转轮从开始转动此后,所有人
就都在命运的流程里生、离、死、别,随着命运之轮的转动永不能再停歇!
十四年以后。
洛阳。朱雀大道。听雪楼。
在堂中所有人七嘴八舌的议论声中,主人冷冷地扬了一下眉毛,然后一抬手——“
唰!”如同一道亮丽的闪电般,我一掠而过,牢牢地钉入檀木茶几。知道主人是要镇住
楼中不服她的人们,于是我尽情地展现着自己的光辉,轻轻摇曳,幻出清影万千。
“血薇剑!”
我一如既往地听见了人们的惊呼,还有窃窃私语,但是,没有人再敢怀疑年轻的主
人的武功和能力——哎,人类都是这样欺软怕硬的吗?看着冷漠美丽的主人,我有些高
兴地笑了。
“你是舒血薇的什么人?”我听见有人惊讶地问主人,看来,前任主人虽然离世那
么多年了,名头依然响亮的很啊……熟悉的手轻轻把我从几上拔起,然后,我听到了主
人淡淡的回答:“——我叫舒靖容,以后叫我阿靖就好。”
堂中的气氛忽然间凝结——我发觉所有人都用很奇怪的目光看着主人,戒备中带着
嫌恶——血魔的女儿——因为这个身份,主人从小受尽了白眼与冷落,没有一个可以说
说话的伙伴,那样孤苦飘零地一个人过了二十二年。
多年过去了,江湖局面也早已经不同往日,然而即使到了现在,居然还是受排斥吗?
从主人八岁起,我就跟着她了……一直到十年后,我和主人才达到了心灵默契的境
地。以后,我能知道她的喜怒哀乐,而她也视我如同她的生命。
她自幼经历的一切,只有我知道,也只有我懂。
那是令任何人都无法忍受的歧视、寂寞、排斥和放逐……但令我安心的是,主人毕
竟没有被打倒,她是那样坚强地活了下来,并且得到了足够在江湖中生存下去、不畏惧
任何人的力量。
但是,经过了那样的童年和少年时期,主人的内心变的惊人的冷漠和孤僻,不依靠
任何人也不相信任何人,拒绝着亲情友情和爱情,唯一相信的,只有力量和命运而已。
——那样苍凉的心境,让我都无论如何不能相信,她还是一个刚刚二十二岁的韶龄
女子。
主人没有说话,修长的手指轻轻抚着我的锋芒,看着面前惊疑的众人,眼睛里有讽
刺的光。
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R*y
2
IPAD一出来就进了, IPAD3只要分辨率高也会进一个. 上网随便看看IPAD还行, 象我开
的网页多, 需要看的东西多, 尤其是照片多(摄影论坛), 经常灌水打字的, 还的笔记本.
IPAD也就随便看看email, 简单上网不错, 玩一些游戏, 打牌, 看网上视频好. 还有就
是看照片好, 但很不喜欢照片得通过ITUNES.
别说我果黑, 我有IPHONE/IPOD.
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x*i
3
Hello there:
Trytophan fluorescence spectroscopy could be used to analyze whether protein
is compact or open. Ex at 288 nm and em around 320 nm suggests that protein
is compact (trytophan in hydrophobic environment). Em at longer wavelength
suggests an open structure. Also relative fluorescence intensity could tell
something? Anybody know why?
Thanks
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w*e
4
It is because the emission wavelength of a fluorophore is very much
influenced by the polarity of the micro-environment. A polar medium makes
the emission "red-shifted", meaning longer wavelength, whereas a non-polar
medium makes the emission blue shifted. Tryptophan is a hydrophobic amino
acid. In a folded protein, it is usually buried within the interior of the
protein (hydrophobic core) so that it is in a relatively non-polar
environment. In a completely unfolded protein, Tryptophan is exposed to
aqueous medium so is in a polar environment. If I remember correctly,
emission at 350nm indicates a completely unfolded protein. The intensity of
a fluorophore is also influenced by polarity, but it is not as useful of a
tool to assess folding and structure as emission wavelength since intensity
is concentration-dependent, and also because intensity is easily subjected
to artifacts such as scattering.
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w*e
5
Part of my master thesis is about fluorescence spectroscopy. Glad to see
that it can be useful.
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x*i
6
Thanks! But why the philic environment causes emission red shift?
Bascially we have a protein soluble at room temperature but it precipitates
at cold temperature such as 4 degree. It re-dissolves when temperature
increases. I am thinking about fluorescence spectroscopy to see compactness
or openness of protein with temperature.
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p*n
7
总体能量变低就红移,变高就蓝移
蛋白不融你怎么做fluorescence?

precipitates
compactness

【在 x*******i 的大作中提到】
: Thanks! But why the philic environment causes emission red shift?
: Bascially we have a protein soluble at room temperature but it precipitates
: at cold temperature such as 4 degree. It re-dissolves when temperature
: increases. I am thinking about fluorescence spectroscopy to see compactness
: or openness of protein with temperature.

avatar
w*e
8
For the first question, that has to do with how fluorescence occurs.
Basically, when excited electrons return to the ground state, some
energy is lost (as heat), and some energy is emitted as photons
(fluorescence). In polar environment, more energy is lost and less is
emitted. For the second question, I think you should look into
differential scanning calorimetry (or other types of calorimetry). That is
a more common technique to study protein folding/unfolding with
temperature changes, which seems to be what you are trying to study.

precipitates
compactness

【在 x*******i 的大作中提到】
: Thanks! But why the philic environment causes emission red shift?
: Bascially we have a protein soluble at room temperature but it precipitates
: at cold temperature such as 4 degree. It re-dissolves when temperature
: increases. I am thinking about fluorescence spectroscopy to see compactness
: or openness of protein with temperature.

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