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s****9
发帖数: 932 | 2 My two cents:
1. Not sure what kind of particle it is. But for lyposome, MPLA can bind
lyposome particles in the outside domains.
2. Still in debate whether TLR4 can be internalized and remain active in the
endosome. But it can be a possibility.
面？ |
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c**g
发帖数: 14 | 3 Unlike TLR3 and TLR7/8/9 which are located and act
exclusively from endosome, TLR4 can signal from both cell surface and
locations within the cell. The difference is that, the activation of TLR4 at
cell surface involves Myd88 and leads to NF-kB pathway activation, while
the activation of TLR4 in endosome involves TRIF and leads to IFN production.
面？ |
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L****S
发帖数: 89 | 4 正解。看前面的回复真是隔行如隔山阿
at
production. |
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S****r
发帖数: 982 | 5 Then, how could it be explained that dsRNA analogy in culture medium
activates TLR3, while transfected dsRNA analogy activates RIG-1/MDA5?
Thanks in advance!
at
production. |
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S****r
发帖数: 982 | 6 Thanks! endosome vs cytosolic, it nicely addressed the question tangled me
long time. |
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c**g
发帖数: 14 | 7 I think your explanation of location is logical, however I have read a paper
which said that the DOTAP transfection reagents (liposome based) can
directly deliver DNA/RNA into the endosome, and the TLR3 located mainly in
the endosome,so why is RIG-1/MDA5 activated when dsRNA is transfected
through liposome? |
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S****r
发帖数: 982 | 8 Does DOTAP bring DNA/RNA contents only to endosome?
paper |
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L****S
发帖数: 89 | 9 Thanks for the discussion. I know which papers you are talking about. We
have to think about immunology here.
In the endosomal compartment, for dsRNA, ssRNA, DNA, TLR3/7/9 sense them
accordingly. No big question here.
In the cytosol, dsRNA is a strong "danger" signal showing viral infection
. That's why every cell has PKR, RIG-I/MDA-5, etc. So I speculate, when
liposome delivers ssRNA, RIG-I/MDA-5-mediated cytosolic response dominates
the TLR3 signal, quantitatively or qualitatively.
In the pape... 阅读全帖 |
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h*****t
发帖数: 103 | 10 为什么不贴出原文呢？
这样类似的文章很多么？是哪里发的呢？
我觉得有时候还是需要考虑本身文章的质量和可信度，不然讨论半天，也许本身实验就
有问题或者实验太超前，一般人理解不了。
另外，活化表面的TLR4之后，也会引起Type I IFN的产生吧？
at
production. |
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i****g
发帖数: 3896 | 11 引用200次以上的4篇：
Title: Directional guidance of neuronal migration in the olfactory system by
the protein Slit
Author(s): Wu W; Wong K; Chen JH; et al.
Source: NATURE Volume: 400 Issue: 6742 Pages: 331-336 Published: JUL
22 1999
Times Cited: 319 (from Web of Science)
Title: Vertebrate slit, a secreted ligand for the transmembrane protein
roundabout, is a repellent for olfactory bulb axons
Author(s): Li HS; Chen JH; Wu W; et al.
Source: CELL Volume: 96 Issue: 6 Pages: 807-818 DOI: 10.10... 阅读全帖 |
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c**n
发帖数: 73 | 12 Cardiovasc Hematol Agents Med Chem. 2011 Jan;9(1):42-55.
CD36 as a multiple-ligand signaling receptor in atherothrombosis.
Nergiz-Unal R, Rademakers T, Cosemans JM, Heemskerk JW.
Source
Department of Biochemistry, Cardiovascular Research Institute Maastricht (CA
RIM), Maastricht University, The Netherlands.
Please send it to c********[email protected]
Thanks a lot in advance. |
|
p********i
发帖数: 116 | 13 请教一个问题，如果要确定一个膜蛋白是一个东西的受体，最常用的实验是什么？谢谢
。 |
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p****s
发帖数: 3153 | 14 不知道最常用的是什么...不过应该可以同位素标记配体然后跑胶吧 |
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G******O
发帖数: 189 | 15 1，表达定位在膜上
2，体外亲和力试验
3，在表达的细胞上用配体处理有特异性信号
4，最终一锤定音的是搞清结构 |
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f********s
发帖数: 115 | 16 想请教一个实验问题：有没有人尝试过用液体培养基加细胞因子来分化过细胞？
我想做这样一个实验，把MEP用FACS分选出来，然后在每一个96孔板的孔里面放一个细
胞。分化7天以后把这个孔里的细胞cytospin + staining， 然后看每个MEP都能分化成
什么种类的子细胞。用这个实验来比较wt 和mut 的区别。我看到一个protocol， 用
IMDM+ SCF，IL3, IL11, GM-SCF, FLT3-Ligand, TPＯ和EPO 来养细胞。我没有做过液
体培养基，一般用的都是methylcult medium,所以不知道液体培养基的效果好不好。有
没有有经验的人来说一说呢？
不知道在哪里看到说semi-solid的培养基比纯液体好，但是考虑到最后要cytospin,
semi-solid的培养基太粘了，洗一下的话又怕细胞丢失太多，因为本来一个细胞就分化
不出太多细胞。
多谢多谢！！ |
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g*****j
发帖数: 1211 | 17 生物在细胞水平的信号传导在动物和植物之间会有很多共通的地方。植物合成的很多
ligand对动物细胞reeceptor 有激活或者抑制作用是完全可能的。比方
catecholamnine。植物中分离的很多物质如tyramine, Digitalis, muscarine，
nicotine，atropine 都可以入药，只是现在有了更好的选择。 中医临床上的阴和阳，
和人体的sympathetic 和 parasympathetic nerve system 的作用有很多重合的地方。
只是古人不知道这些生理机制。 |
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j*****d
发帖数: 787 | 18 我现在操心的是 细胞 怎么把外源的miRNA 内吞到胞内？分泌部分似乎解决了。以后能
够miRNA 作 ligand 就爽了。 |
|
j*****d
发帖数: 787 | 19 我现在操心的是 细胞 怎么把外源的miRNA 内吞到胞内？分泌部分似乎解决了。以后能
够miRNA 作 ligand 就爽了。 |
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g*****n
发帖数: 241 | 20 我能想到的间接证明的方法是 non-allelic non-complimentation test
请问其他有没有什么方法？
谢谢 |
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b****r
发帖数: 17995 | 21 遗传的方法只能证明上下游关系吧，不能证明直接的binding |
|
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y******8
发帖数: 1764 | 23 You have to set a cellular phenotype for direct interaction.
You might call yeast two hybrid as genetics. |
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C*****h
发帖数: 926 | 24 You may want to try some ligands that are specific to T cell receptors to
deliver drugs.
function |
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g****y
发帖数: 101 | 25 To Carwash: Thank you very much for comment
I can't agree more with you about the low efficiency in infecting the T
cells in vivo with virus. That is why I mentioned the potential problem 4(
AAV载体的靶向问题).
As you suggested, we could modify the virus caspid, for example, mutate the
liver cell recognition sequence and express a chimeric protein combining the
AAV capsid protein and the T cell ligand so that we can increase the
infection efficiency in T cells.
Do you think this approach works?
Here ar... 阅读全帖 |
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C*****h
发帖数: 926 | 26 我知道日本有一家公司，他们用一种脂质体，上面经过修饰加工，
带有T cell receptor的ligand，脂质体里面包埋逆转录病毒的RNA。
脂质体把RNA送到T cells里面，再逆转录，整合到genome
the
the |
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g****y
发帖数: 101 | 27 liver accumulation 可能是因为liver有清除外来异物的功能吧， 我打算用的病毒也
是在liver有很高的感染效率，所以我打算突变掉病毒capsid的肝脏识别序列并替换成T
cell ligand， 希望这样能提高感染T细胞的效率， 不知道这种设想可不可行， 赫赫 |
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n****n
发帖数: 610 | 28 有篇paper学校下不了，不知道哪位能否帮忙下一下？我的email: n****[email protected]
包子答谢！
paper title: Nondenaturing Mass Spectrometry to Study Noncovalent Protein/
Protein and Protein/Ligand Complexes: Technical Aspects and Application to
the Determination of Binding Stoichiometries |
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n********k
发帖数: 2818 | 29 This is a smart, very focused and productive one I have heard from people
working with him...XD Wang's first student was even more impressive if I am not too old:) and several from HJ Song's lab also have impressive records, A and P from this board both have impressive records in pretty shoret time too...
BTW, to upbeat/Q a bit for those (including myself) who may feel depressed now)....Nothing particularly/unusually, Pretty much decent intelligence, strong motivations/working/person habits/... 阅读全帖 |
|
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a*****v
发帖数: 128 | 31 大家都是如何拟合滴定的数据呢？
在蛋白质浓度不足的情况下，只能滴加ligand的方式进行，这个时候蛋白质的浓度在不
断稀释，
因此不能用简单的y=B*x/(x+kd) 来拟合
大家是用什么软件，如何做的呢？ |
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t***i
发帖数: 204 | 32 M Azuma. (2010) Role of the Glucocorticoid-Induced TNFR-Related Protein (
GITR)-GITR Ligand Pathway in Innate and Adaptive Immunity. Critical Reviews
in Immunology, 30(6):547-557
Please send to t***********[email protected]. Thanks a lot. |
|
t***i
发帖数: 204 | 33 M Azuma. (2010) Role of the Glucocorticoid-Induced TNFR-Related Protein (
GITR)-GITR Ligand Pathway in Innate and Adaptive Immunity. Critical Reviews
in Immunology, 30(6):547-557
Please send to t***********[email protected]. Thanks a lot. |
|
b******y
发帖数: 627 | 34 I agree with the referee. But does the absolute Kd matter in your case or
the relative Kd? For example, you want to demonstrate wt and mutant proteins
have a big difference in binding the same ligand. In either case, to a
reasonable referee, you might be OK if using Kapp instead of Kd in the paper
and also acknowledge your agreement with his/her opinion in the rebuttal
letter. Good luck! |
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t***i
发帖数: 204 | 35 M Azuma. (2010) Role of the Glucocorticoid-Induced TNFR-Related Protein (
GITR)-GITR Ligand Pathway in Innate and Adaptive Immunity. Critical Reviews
in Immunology, 30(6):547-557
Please send to t***********[email protected]. Thanks a lot. |
|
S***a
发帖数: 257 | 36 pls send to columxia at gmail.com
Thank you and happy new year!
Evaluation of the carbohydrate recognition domain of the bacterial adhesin
FimH: design, synthesis and binding properties of mannoside ligands Oliver
Sperling, Andreas Fuchs and Thisbe K. Lindhorst
Org. Biomol. Chem., 2006, 4, 3913-3922 |
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l*****4
发帖数: 267 | 37 通过什么实验可以在动物体内直接证明 steroid 和不和 steroid receptor 结合？
谢谢 |
|
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l**********1
发帖数: 5204 | 39 RE LD
你这问题问的是高中生高考水平或生化系大学3年前水平的话
答案是 stable同位素标定Steroid Legend 然后 assay 然后单离steroid receptor 质谱分析receptor里边有无
那同位素
如果问的是PD PhD thesis 水平
输入你那个目标steroid receptor 之
Protein ID Protein Sequence
然后
http://www.pathblast.org/
查一下先
偶这也是借以色列的某教授的花
http://www.cs.tau.ac.il/~roded/
Selected Web-Servers / Software:
· ANAT
· Torque
· NetworkBLAST
· EXPANDER
献佛仅此而已 |
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l**********1
发帖数: 5204 | 40 Two ways:
No. 1
Patch clamp Single-channel recording
htp://parkerlab.bio.uci.edu/publication%20attachments/Demuro2003_CellCalcium_120.pdf
No. 2
single-molecule fluorescence methods
hp://iopscience.iop.org/1478-3975/7/3/031001/pdf/1478-3975_7_3_031001.pdf
have the possibilities i think. |
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l**********1
发帖数: 5204 | 41 Re rien
three BAOZI got, Thank you.
BTW, plus one paper:
Predicting protein ligand binding motions with the Conformation Explorer
Flores, S.C. and Gerstein, M.B. (2011) BMC Bioinformatics, 12: 417 (Highly
Accessed).
//www.ncbi.nlm.nih.gov/pubmed/22032721
its corresponding author link:
//xray.bmc.uu.se/flores/Papers.html |
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K**4
发帖数: 1015 | 42 做点突变 ，测酶活性而已
不需要什么 计算
况且结构计算 已经没有人 相信了
连文章 都看不到了
往往蛋白质活性中心有 一组residues用于催化
同时还有 几个 residues用于 bind ligand.
所以你只要根据sequence／structure找到 这些 residues，做个突变就可以吧 |
|
z****u
发帖数: 1007 | 43 正想问同样问题。这几天在做调查。
好像这个显微镜能看到ligand-receptor binding那个层次。
这显微镜要多少钱？ 比如说Leica SR GSD. |
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z****u
发帖数: 1007 | 44 正想问同样问题。这几天在做调查。
好像这个显微镜能看到ligand-receptor binding那个层次。
这显微镜要多少钱？ 比如说Leica SR GSD. |
|
C*******e
发帖数: 4348 | 45 谢谢回复
我用的是1ml的ITC
只有第一个full volume injection看到比background高一点的峰
一个是蛋白浓度低
还有一个可能一下就饱和了（ligand:protein=24:1)
现在准备用更高的蛋白浓度看看
如果第一个峰更高了
可能会再做一次 |
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i*******e
发帖数: 62 | 46 请教各位一个关于线性范围的问题。
大家都知道酶反应动力学中的Michaelis–Menten方程（米氏方程）。许多ligand-
receptor binding反应也符合这一方程。这种反应的特征性曲线就是hyperbolic curve
.在尚未达到saturation (plateau phase)之前，有一段曲线近似于直线。位于这一线
性范围内（linear range）的数据有着近乎linear regression （R2>0.99）的关系。
请教如何可以比较准确的确定这一线性范围。有没有相关文献可以参考？万分感谢！ |
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l**********1
发帖数: 5204 | 47 那棒子吗? HT 还力压Zhuang XW 呢 那会那篇Steven Chu NAS PNAS contribution
paper:
Ligand-induced conformational changes observed in single RNA molecules
Contributed by Steven Chu, May 27, 1999
//www.ncbi.nlm.nih.gov/pubmed/10430898 |
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g*********9
发帖数: 3528 | 48 It is clearly stated on its website......
See background:
"Stat3 isoform expression appears to reflect biological function as the
relative expression levels of Stat3α (86 kDa) and Stat3β (79 kDa) depend
on cell type, ligand exposure, or cell maturation stage (10). It is notable
that Stat3β lacks the serine phosphorylation site within the carboxy-
terminal transcriptional activation domain (8)." |
|
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p*****u
发帖数: 276 | 50 itc能否用，还要看你的结合常数大小，如果结合常数太弱就不行，还有你的蛋白质浓
度也很重要，还要保证蛋白质不变性，另外滴定的那个ligand不能太粘否则剂量就不准
了。你的这个情况基本不能用itc，可以考虑spr（表面质子共振），快速便宜还方便。
就是在chip上连接你的蛋白质，然后看激光的折射率变化，如果有binding那折射率就
变化了，根据变化可以计算出结合常数。 |
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