l**********1
发帖数: 5204 | 1 //www.ncbi.nlm.nih.gov/pubmed/20023165
full text link:
//www.weigelworld.org/research/publications/2010/pdfs/Rodriguez%
20Development%202010.pdf
even Xue-mei Chen before 2010 never did Arabidopsis cyclin-dependent
kinases and microRNA
after 2011 she should noted that paper.
And You centuribob (Planta) from 2012 now still ignored it? |
|
l**********1
发帖数: 5204 | 2 你就差多玛三个字
的关联=XYZ below shown
Xuemei又不做抗性和kinase（XYZ) ，人家做小RNA的代谢
害得我还以为你是她的原同事呢
BTW
既然centuribob (Planta) 的大 是专搞RNA world的
是Yonath Ada 的lab出来的薄厚 /PI 你认识吗? i mean met him/her before whom was PD
from her:
//www.nobelprize.org/nobel_prizes/chemistry/laureates/2009/
//www.weizmann.ac.il/sb/faculty_pages/Yonath/
Weizmann Institute of Science, Rehovot, IL,
The amazing ribosome
link:
//www.euchems-prague2012.cz/files/euchems-adv-210x279-05.pdf |
|
a*****x
发帖数: 901 | 3 不是很理解，Acetyltransferases为什么是Kinases的counterparts,都不是作用同样的
residue。。。 |
|
y*******a
发帖数: 303 | 4
我们也是用高保真酶P的全长。我是觉得colony PCR 没必要用高保真酶检验。我的
promoter是35S，不知道是不是E.Coli体内无法接受protein kinase? |
|
y*******a
发帖数: 303 | 5
我可以得到基因的全长，只是觉得用高保真酶做colony PCR比较浪费。我用的promoter
是35S.不知道是不是因为E.Coli自身接受不了protein kinase的表达？ |
|
o**d
发帖数: 3080 | 6 有点不太明白了。
你是要用ecoli来表达kinase这个protein还是只有用ecoli来做克隆？
promoter |
|
y***y
发帖数: 709 | 7 有一个蛋白，预测到多个磷酸化位点
想看看都什么激酶作用这些位点，不知道kinase profiling是不是能达到这个目的。
看到很多公司有这个sevice，不知板上是否有人用过这个sevice，效果怎么样？那个公
司比较靠谱？大概收费是多少？ |
|
p*****e
发帖数: 93 | 8 put in GPS 2.1 (a free software), it will tell you the potential kinases. |
|
y***y
发帖数: 709 | 9 我就是用这个软件预测的磷酸化位点。
可预测出来的潜在位点太多了，我想通过Kinase profiling的方法把作用于这个蛋白的
激酶筛出来。 |
|
S*********s
发帖数: 304 | 10 我觉得你这个jump太多了
至少要先确定是哪个点被phosphorylated了吧？
是不是functional的？
www.phosphosite.org里面可以查查MS data
In vitro kinase profiling我总觉得很不特异 |
|
y***y
发帖数: 709 | 11 您可以稍微解释一下您说的in vivo crosslink吗？
我们已经做了MS，并确认了几个位点确实有磷酸化
我坐在这里想象一下：
如果有这么一种slide，上面点着各种不同激酶的特异性抗体，当用激酶的混合物和
slide杂交之后，激酶就会结合在各自的抗体之上，洗掉激酶混合物以及非特异性结合
之后，放在目标蛋白和标记了的ATP溶液中，一段时间后加蛋白交联剂，把和激酶作用
的目标蛋白交联在激酶和抗体复合物上，洗脱，扫片子，有标记信号的点表示该抗体所
对应的激酶们就是目标蛋白磷酸化的激酶。
这个实验能够实现的话，那就是满足我需要的dream experiment。其中最难的应该是蛋
白交联这一步了。如果有公司想研发这种真正的kinase profiling的话，尽管拿这个
idea去研发，我不收专利费，哈哈 |
|
b****u
发帖数: 2771 | 12 curious question: what is time scale for kinase and substrate binding. |
|
p*****e
发帖数: 93 | 13 Check whether the phosphorylation sites match the phosphorylation motif of
these potential kinases. |
|
S*********s
发帖数: 304 | 14 many kinase/substrate interaction is transient
can not get interaction by IP unless using crosslinker |
|
l****y
发帖数: 398 | 15 but the kinases are still active. i want them all dead. |
|
s******n
发帖数: 110 | 16 激酶检测（kinase assay）是在试管中进行的（in vitro），每次只有一个底物，要么
是A，要么是B，而且都是纯化的蛋白。因此它们之间的影响被消除了。 |
|
b******y
发帖数: 627 | 17 What emases meant is that A or B or both might allosterically regulate ATP'
s binding affinity to kinase and therefore the Km of ATP can be different
when A or B as substrate due to differential allosteric effects from A and B
. |
|
g******o
发帖数: 96 | 18 target protein可以被上游kinase磷酸化。serine突变成aspartic acid有表型，可是
serine突变成alanine没有表型。reviewer认为这个位点在正常生理情况下不怎么重要
，并要求解释原因。我的解释是这个位点在in vivo 被高度调控，可能有phosphate不
停的去磷酸化；也可能只有在特定的条件下才被磷酸化。
有人知道有这样的例子吗？只有D mutation 有表型而A mutation 没有表型的。
谢谢！ |
|
p*****c
发帖数: 116 | 19 跟你用的细胞株有关，有些肿瘤细胞内kinase活性超强，WT transfection其实自动被
磷酸化变成D mut状态。 |
|
p*****h
发帖数: 36 | 20 Raf has very few known substrates (MEK)
For the majority, specificity is determined by the activation segment of the
kinase domain. |
|
p*****h
发帖数: 36 | 21 You mean not just MEK?
Raf does not have a specificity motif. Peptide library approaches give no
definitive results. - this is generally taken as evidence that sequences
outside the kinase domain help anchor substrates. |
|
p*******1
发帖数: 495 | 22 做一个Ser/Thr kinase激酶活性的检测，它可以自体磷酸化，想问一下大家，除了同位
素P32外，还有什么办法可以检测其激酶的活性，有没有什么非同位素的试剂盒可以使
用？
谢谢了先！ |
|
V*******s
发帖数: 19 | 23 Does anyone know the companies that have products for Kinase arrays?
Thanks! |
|
m****g
发帖数: 530 | 24 invitrogen, the they provide comprehensive assays tools and services. the
other one is Ambit bioscience. they provide kinome wide kinase screening. |
|
l****y
发帖数: 486 | 25 Not sure about your point here.
For example, if you are studying a kinase function in neurobiology. You
found total KO of your gene leads to embryonic lethal phenotype, and
conditional KO of your gene in neuron leads to very interesting phenotype.
Culturing of KO neuron in vitro also showed dramatic phenotype.
Interestingly, restoration of kinase dead (KD) mutant in KO neuron fully
rescued the phenotype (same as restoration of WT).
You wonder whether the neuronal phenotype you observed in vivo i... 阅读全帖 |
|
A********e
发帖数: 354 | 26 【 以下文字转载自 Biology 讨论区 】
发信人: albertsmwk (.)(.), 信区: Biology
标 题: 第一批“青年千人计划”生物类@publication列表@
发信站: BBS 未名空间站 (Thu Aug 18 15:04:16 2011, 美东)
花了一个小时，深深的鄙视一下自己的无聊行径！！
125 蔡亮 男 1980年11月 复旦大学 生命
科学 2007年12月毕业于[美国]北卡大学 [美国]加州大学旧金山分校 博士后
Cai L, Mostov K. Polarity is destiny. Cell. 2009 Nov 13;139(4):660-2. PubMed
PMID: 19914162; PubMed Central PMCID: PMC2900917.
Cai L, Makhov AM, Schafer DA, Bear JE. Coronin 1B antagonizes cortactin and
remodels Arp2/3-containing actin branche... 阅读全帖 |
|
p*******g
发帖数: 2976 | 27 【 以下文字转载自 Biology 讨论区 】
发信人: albertsmwk (.)(.), 信区: Biology
标 题: 第一批“青年千人计划”生物类@publication列表@
发信站: BBS 未名空间站 (Thu Aug 18 15:04:16 2011, 美东)
花了一个小时，深深的鄙视一下自己的无聊行径！！
125 蔡亮 男 1980年11月 复旦大学 生命
科学 2007年12月毕业于[美国]北卡大学 [美国]加州大学旧金山分校 博士后
Cai L, Mostov K. Polarity is destiny. Cell. 2009 Nov 13;139(4):660-2. PubMed
PMID: 19914162; PubMed Central PMCID: PMC2900917.
Cai L, Makhov AM, Schafer DA, Bear JE. Coronin 1B antagonizes cortactin and
remodels Arp2/3-containing actin branche... 阅读全帖 |
|
O***n
发帖数: 13127 | 28 【 以下文字转载自 Biology 讨论区 】
发信人: albertsmwk (.)(.), 信区: Biology
标 题: 第一批“青年千人计划”生物类@publication列表@
发信站: BBS 未名空间站 (Thu Aug 18 15:04:16 2011, 美东)
花了一个小时，深深的鄙视一下自己的无聊行径！！
125 蔡亮 男 1980年11月 复旦大学 生命
科学 2007年12月毕业于[美国]北卡大学 [美国]加州大学旧金山分校 博士后
Cai L, Mostov K. Polarity is destiny. Cell. 2009 Nov 13;139(4):660-2. PubMed
PMID: 19914162; PubMed Central PMCID: PMC2900917.
Cai L, Makhov AM, Schafer DA, Bear JE. Coronin 1B antagonizes cortactin and
remodels Arp2/3-containing actin branche... 阅读全帖 |
|
a********k
发帖数: 2273 | 29 花了一个小时，深深的鄙视一下自己的无聊行径！！
125 蔡亮 男 1980年11月 复旦大学 生命
科学 2007年12月毕业于[美国]北卡大学 [美国]加州大学旧金山分校 博士后
Cai L, Mostov K. Polarity is destiny. Cell. 2009 Nov 13;139(4):660-2. PubMed
PMID: 19914162; PubMed Central PMCID: PMC2900917.
Cai L, Makhov AM, Schafer DA, Bear JE. Coronin 1B antagonizes cortactin and
remodels Arp2/3-containing actin branches in lamellipodia. Cell. 2008 Sep
5;134(5):828-42. PubMed PMID: 18775315; PubMed Central PMCID: PMC2570342.
Cai L, Makhov AM, Bear JE. F-actin... 阅读全帖 |
|
j*b
发帖数: 341 | 30 这个PKM2和肿瘤代谢的关系已经知道很久了，这两篇还不是原创贴。
cantley还可以发nature，能不能解释一下？
Anticancer Res. 2003 Mar-Apr;23(2A):1155-8.
Tumor marker pyruvate kinase type tumor M2 in patients suffering from
diabetic nephropathy.
Oremek GM, Rutner F, Sapoutzis N, Sauer-Eppel H.
Source
Central Laboratory, Hospital of the Johann Wolfgang Goethe-University,
Frankfurt/Main, Germany.
Abstract
INTRODUCTION:
Compared to normal tissues all tumors investigated so far and especially
those with a metastatic potential overexpress an isoenzy... 阅读全帖 |
|
j*b
发帖数: 341 | 31 这个PKM2和肿瘤代谢的关系已经知道很久了，这两篇还不是原创贴。
cantley还可以发nature，能不能解释一下？
Anticancer Res. 2003 Mar-Apr;23(2A):1155-8.
Tumor marker pyruvate kinase type tumor M2 in patients suffering from
diabetic nephropathy.
Oremek GM, Rutner F, Sapoutzis N, Sauer-Eppel H.
Source
Central Laboratory, Hospital of the Johann Wolfgang Goethe-University,
Frankfurt/Main, Germany.
Abstract
INTRODUCTION:
Compared to normal tissues all tumors investigated so far and especially
those with a metastatic potential overexpress an isoenzy... 阅读全帖 |
|
p*******g
发帖数: 2976 | 32 【 以下文字转载自 Biology 讨论区 】
发信人: albertsmwk (.)(.), 信区: Biology
标 题: 第一批“青年千人计划”生物类@publication列表@
发信站: BBS 未名空间站 (Thu Aug 18 15:04:16 2011, 美东)
花了一个小时，深深的鄙视一下自己的无聊行径！！
125 蔡亮 男 1980年11月 复旦大学 生命
科学 2007年12月毕业于[美国]北卡大学 [美国]加州大学旧金山分校 博士后
Cai L, Mostov K. Polarity is destiny. Cell. 2009 Nov 13;139(4):660-2. PubMed
PMID: 19914162; PubMed Central PMCID: PMC2900917.
Cai L, Makhov AM, Schafer DA, Bear JE. Coronin 1B antagonizes cortactin and
remodels Arp2/3-containing actin branche... 阅读全帖 |
|
c**i
发帖数: 6973 | 33 Peter Landers, New Breast Cancer Drug Found Deep in the Sea. Wall Street
Journal, Jan. 4, 2010.
http://online.wsj.com/article/SB1000142405274870411150457605977
("A study found it [Halaven] extended the life of patients with advanced
breast cancer by about two-and-1/2 months, to 13 months")
My comment:
(a) Eisai Co., Ltd. エーザイ株式会社
(a public company; headquarters Tokyo; established in 1941 by Toyoji NAITO
内藤 豊次 as Nohon Eisai Co., Ltd. 日本衛材（衛生材料の略。具体的には絆創膏
や包帯の事))
ja.wikipedia.org
(b) eribulin
htt... 阅读全帖 |
|
c**i
发帖数: 6973 | 34 Peter Landers, New Breast Cancer Drug Found Deep in the Sea. Wall Street
Journal, Jan. 4, 2010.
http://online.wsj.com/article/SB1000142405274870411150457605977
("A study found it [Halaven] extended the life of patients with advanced
breast cancer by about two-and-1/2 months, to 13 months")
My comment:
(a) Eisai Co., Ltd. エーザイ株式会社
(a public company; headquarters Tokyo; established in 1941 by Toyoji NAITO
内藤 豊次 as Nohon Eisai Co., Ltd. 日本衛材（衛生材料の略。具体的には絆創膏
や包帯の事))
ja.wikipedia.org
(b) eribulin
htt... 阅读全帖 |
|
w***7
发帖数: 1637 | 35 source：http://bbs.netbig.com/thread-2491712-1-2.html
看了一下，今年不错啊，和清华并驾齐驱。
而且下半年我知道的还有两篇NS。
~~~~~~~~~~~~~~~~~
1.
Yuan W, Wu T, Fu H, Dai C, Wu H, Liu N, Li X, Xu M, Zhang Z, Niu T, Han Z,
Chai J, Zhou XJ, Gao S, Zhu B
Dense Chromatin Activates Polycomb Repressive Complex 2 to Regulate H3
Lysine 27 Methylation
Science, DOI: 10.1126/science.1225237, Aug. 24, 2012
2.
Liu B, Du H, Rutkowski R, Gartner A, Wang X
LAAT-1 Is the Lysosomal Lysine/Arginine Transporter That Maintains Amino
Acid Homeostasis
S... 阅读全帖 |
|
S*********s
发帖数: 304 | 36 最基本的一条就是癌细胞harbors a lot of mutations,
这些mutant蛋白需要正确folding的话需要chaperon来辅助。
例如，lots of cancers have kinase mutation or overexpression.
这些kinase需要hsp来辅助他们folding.
Neal Rosen同志一直在推Hsp90 inhibitor的clinical trial,
但目前为止没有很好地结果，(GARAMYCIN,17-AAG等一系列衍生物)
其实很多东西都能抑制Hsp90,植物及中药中的很多terpinoid家族化合物都可以。
中药不能乱吃啊~~刚刚三期被terminated kidney failure drug RTA402本身就是
Hsp90 inhibitor,虽然一二期骗了很多钱，但最终还是以失败而告终。
业界及学界都不看好Hsp90 inihibitors,原因主要有二：
一方面是由于是没有特异性，后期toxicity会很难办。
另一方面，由于Hsp90/cdc37 complex是kinase的chaperon,
所以... 阅读全帖 |
|
c**i
发帖数: 6973 | 37 Peter Landers, New Breast Cancer Drug Found Deep in the Sea. Wall Street
Journal, Jan. 4, 2010.
http://online.wsj.com/article/SB1000142405274870411150457605977
("A study found it [Halaven] extended the life of patients with advanced
breast cancer by about two-and-1/2 months, to 13 months")
My comment:
(a) Eisai Co., Ltd. エーザイ株式会社
(a public company; headquarters Tokyo; established in 1941 by Toyoji NAITO
内藤 豊次 as Nohon Eisai Co., Ltd. 日本衛材（衛生材料の略。具体的には絆創膏
や包帯の事))
ja.wikipedia.org
(b) eribulin
htt... 阅读全帖 |
|
p***o
发帖数: 714 | 38 【 以下文字转载自 Stockcafeteria 俱乐部 】
发信人: jhsph07 (银杏), 信区: Stockcafeteria
标 题: Re: ZT: EXEL
发信站: BBS 未名空间站 (Mon Jul 6 11:51:53 2009, 美东)
基本的意思是说， EXEL是做KI的， KI的成功率大概是４０％左右， EXEL的pipeline
上面大概有11个KI， 如果按统计规律， 应该有5个最后会被FDA
批准。
等果实最终成熟大概还需要两年的时间。
Exelixis proves skeptics wrong
Dec 19, 2008
Another company that gives its shareholders reasons to smile is Exelixis, following the mammoth deal with BMS. Exelixis licensed its most advanced compound, XL184, and its early stage RAF inhibitor, XL281 in a de... 阅读全帖 |
|
s**u
发帖数: 9035 | 39 He has 12 papers, 3 of which are first-author papers. see below
Differential regulation of androgen receptor by PIM-1 kinases via
phosphorylation-dependent recruitment of distinct ubiquitin E3 ligases.
Linn DE, Yang X, Xie Y, Alfano A, Deshmukh D, Wang X, Shimelis H, Chen H, Li
W, Xu K, Chen M, Qiu Y.
J Biol Chem. 2012 Jun 29;287(27):22959-68. Epub 2012 May 14.
PMID:
22584579
[PubMed - indexed for MEDLINE]
Related citations
2.
A Role for OCT4 in Tumor Initiation of Drug-Resistant Prostat... 阅读全帖 |
|
s**u
发帖数: 9035 | 40 Dr.Xie has 12 papers, 3 of which are first-author papers. see below
Differential regulation of androgen receptor by PIM-1 kinases via
phosphorylation-dependent recruitment of distinct ubiquitin E3 ligases.
Linn DE, Yang X, Xie Y, Alfano A, Deshmukh D, Wang X, Shimelis H, Chen H, Li
W, Xu K, Chen M, Qiu Y.
J Biol Chem. 2012 Jun 29;287(27):22959-68. Epub 2012 May 14.
PMID:
22584579
[PubMed - indexed for MEDLINE]
Related citations
2.
A Role for OCT4 in Tumor Initiation of Drug-Resistant Pro... 阅读全帖 |
|
s******9
发帖数: 283 | 41 NCBI protein database一般都有标注吧。
FEATURES Location/Qualifiers
source 1..503
/organism="Homo sapiens"
/db_xref="taxon:9606"
gene 1..503
/gene="TGFBR1"
/gene_synonym="ALK5"
/gene_synonym="SKR4"
Protein 1..503
/gene="TGFBR1"
/gene_synonym="ALK5"
/gene_synonym="SKR4"
... 阅读全帖 |
|
l*****7
发帖数: 8463 | 42 Cancer researcher retracts 19 studies at once
with 23 comments
jcheng
Jin Cheng
A former cancer biologist at the Moffitt Cancer Center in Tampa, Florida has
retracted 19 papers from a single journal.
Jin Cheng, who studies how ovarian cancer develops, withdrew 19 papers from
the Journal of Biological Chemistry originally published over the last 15
years, and corrected another. All of the retractions are for image
manipulation.
For example, here’s the notice for “Activation of phosphatidylinosito... 阅读全帖 |
|
s*****g
发帖数: 7857 | 43 http://en.wikipedia.org/wiki/Imatinib
tyrosine-kinase inhibitor
is a tyrosine-kinase inhibitor used in the treatment of multiple cancers,
most notably Philadelphia chromosome-positive (Ph+) chronic myelogenous
leukemia (CML).[1] Like all tyrosine-kinase inhibitors, imatinib works by
preventing a tyrosine kinase enzyme, in this case BCR-Abl, from
phosphorylating subsequent proteins and initiating the signaling cascade
necessary for cancer development, thus preventing the growth of cancer cells
an... 阅读全帖 |
|
X******n
发帖数: 914 | 44 Lack of Evidence for PKM2 Protein Kinase Activity－Molecular Cell
Highlights
•PEP-dependent phosphorylation was assayed in cell lysates using
radioactive PEP
•Trace adenine nucleotides can confound PEP-dependent phosphorylation
assay results
•PEP-dependent phosphorylation is rare and is not dependent on PKM2
•We are unable to detect PKM2 protein kinase activity in a variety of
contexts
Summary
The role of pyruvate kinase M2 (PKM2) in cell proliferation is controversial
. A... 阅读全帖 |
|
w***a
发帖数: 4361 | 45 俺正好用过这个pGEX-2TK, 它这个Kinase recognition site不是酶切用的，Thrombin的
切点在旁边的位置。
如果原核表达出来的蛋白，需要同位素标记的话，可以利用这个Kinase识别位点做in v
itro kinase assay。如果你表达GST-fusion蛋白的目的是为了检测这个蛋白是否能被某
个kinase磷酸化，那就不建议用这个载体，否则就无所谓。因为用Thrombin可以把所有
的无关序列都给切掉。 |
|
b****n
发帖数: 311 | 46 老大，你鸣不平也要做点homework吧？
郭岩怎么着都有5篇plant cell. 就算他去年review,去掉2010年的2篇, 那也有3篇
plant cell.
1: Zhou Y, Zhang J, Lin H, Guo G, Guo Y. MORPHEUS' MOLECULE1 is required to
prevent aberrant RNA transcriptional read-through in Arabidopsis. Plant
Physiol.
2010 Nov;154(3):1272-80. Epub 2010 Sep 8. PubMed PMID: 20826701; PubMed
Central
PMCID: PMC2971605.
2: Zhang C, Guo H, Zhang J, Guo G, Schumaker KS, Guo Y. Arabidopsis cockayne
syndrome A-like proteins 1A and 1B form a complex with CULLIN4 and damage
DNA
bind... 阅读全帖 |
|
k**n
发帖数: 6198 | 47 I am not an expert but happened to be in a seminar recently. Many other
kinases, like adenosine monophosphate-activated protein kinase, Jun kinase,
extracullular signal-related kinase ETC. |
|
g********0
发帖数: 6201 | 48 Insubordination in the Lab
05/23/2012 David Pittman
In two recent cases, post-docs have published data without their principal
investigator’s consent. Can this be prevented from happening again?
Jiasheng Diao was a post-doc at the Purdue University, working in the lab of
X-ray crystallographer Miriam Hasson on structure of the protein butyrate
kinase when Hasson passed away in 2006. As a result, the university gave
control of her protein structure data to her husband, Purdue biochemist
David San... 阅读全帖 |
|
K**4
发帖数: 1015 | 49 I didnt read the paper but from what you have said, it is impossible that
small molecule kinase can catalyze proteins.If it is indeed a small molecule
kinase, and his paper showed it is protein kinase, I think he definitely
manipulated the data.
Sorry I dont have Chinese input right now.
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