BIOCHEMICAL RESEARCH ON OPIOID RECEPTORS
BIOCHEMICAL RESEARCH ON OPIOID RECEPTORS
批准号:
6634123
负责人:
Eric J Simon
金额:
$12.71万
依托单位国家:
美国
项目类别:
财政年份:
1999
资助国家:
美国
项目状态:
已结题
起止时间:
1999-05-01 至 2004-02-29
关键词:
G protein biological signal transduction cytoskeleton electrospray ionization mass spectrometry intermolecular interaction matrix assisted laser desorption ionization opioid receptor posttranslational modifications protein purification protein structure function receptor binding receptor expression structural biology
中文摘要
描述(申请人摘要):
这是一份高级科学家奖(K05)的申请书。 的
申请人计划继续他长期的研究,
长期目标是阐明急性和慢性
鸦片的影响。为此,我们正在研究内源性阿片样物质
系统,我们的研究重点是结构,功能,
调节不同类型的阿片受体。 的理解
这种内源性神经肽/受体系统最终应该有
对治疗和预防药物滥用的重要影响
改善了疼痛治疗。
第一个目标涉及活性μ阿片样物质的结构研究
受体蛋白(ORP)。 翻译后
纯化的μ受体的修饰,如糖基化,
磷酸化和二硫桥,将由强大的研究
与罗恩比维斯博士合作,
这项技术的顶尖专家 这种方法也将是有益的
用于探索其他修饰,例如C-
末端半胱氨酸和通过巯基试剂的半胱氨酸烷基化。
在早期制剂中存在的蛋白质,可产生缩氨酸
在牛μ阿片受体序列中没有发现,
最近克隆的,也不在任何数据库中,将被纯化,
其特征在于,因为它可能是一种重要的,新的μ调节剂,
受体功能
半胱氨酸残基在受体功能中的作用将通过
定点诱变和亲和标记。 重点将放在
参与配体结合的残基和C-末端中可能
被棕榈酰化。 脂肪酰化的作用将通过
比较非棕榈酰化突变体与野生型的功能
型受体,并通过研究激动剂和拮抗剂的作用
和其他受体调节剂对动态棕榈酰化速率的影响
受体。 替代半胱氨酸可及性方法(SCAM)将
用于探测阿片样物质的亲水性进入缝隙
受体结合位点 简言之,结合位点中或附近的氨基酸
将被半胱氨酸取代。 配体结合失活,
亲水性SH试剂被认为是半胱氨酸面对
亲水性通道裂缝。
第二个目标是探索阿片受体与
细胞骨架元素和连接受体的蛋白质的存在
到细胞骨架。 这样的相互作用对于正确的
受体的位置和方向,甚至调节
受体功能在最后的目标,信号转导方面,
将进行研究,包括识别涉及的受体区域
在G蛋白激活中。
英文摘要
DESCRIPTION (Applicant's Abstract):
This is an application for a senior scientist award (K05). The
applicant plans to continue his long standing research with the long
term goal of elucidating the molecular basis of the acute and chronic
effects of opiates. To this end we are studying the endogenous opioid
system and the focus of our research is on the structure, function, and
regulation of the different types of opioid receptors. An understanding
of this endogenous neuropeptide/receptor system should ultimately have
important implications for the treatment and prevention of drug abuse
and improved treatment of pain.
The first aim involves structural studies of the active mu opioid
receptor protein (ORP) we have purified. Posttranslational
modifications of the purified mu receptor, such as glycosylation,
phosphorylation and disulfide bridges, will be studied by the powerful
technique of mass spectrometry, in collaboration with Dr. Ron Beavis,
a leading expert in this technology. This approach will also be useful
for the exploration of other modifications such as palmitoylation of C-
terminal cysteines and alkylation of cysteines by sulfhydryl reagents.
The protein, present in earlier preparations, that gave rise to peptides
not found in the sequence of the bovine mu opioid receptor we have
recently cloned, nor in any database, will be purified and
characterized, since it may be an important, novel regulator of mu
receptor function.
The role of cysteine residues in receptor function will be studied by
site-directed mutagenesis and affinity labeling. The focus will be on
residues involved in ligand binding and those in the C-terminal that may
be palmitoylated. The role of fatty acylation will be examined by
comparing the functions of non-palmitoylated mutants with those of wild
type receptors and by studying the effects of agonists and antagonists
and other receptor regulators on the dynamic palmitoylation rate of
receptors. The Substituted Cysteine Accessibility Method (SCAM) will
be employed to probe the hydrophilic access crevice of the opioid
receptor binding site. Briefly, amino acids in or near the binding site
will be replaced by cysteines. Inactivation of ligand binding by
hydrophilic SH reagents is taken as evidence that the cysteine faces the
hydrophilic access cleft.
The second aim proposed probing the interaction of opioid receptors with
cytoskeletal elements and the existence of proteins that link receptors
to the cytoskeleton. Such an interaction may be essential for correct
receptor location and orientation and even for the regulation of
receptor functions. In the final aim, aspects of signal transduction
will be studied, including identification of receptor regions involved
in G protein activation.
期刊论文(2)
专著(0)
科研奖励(0)
会议论文
DOI:
10.1186/1471-2210-2-5
发表时间:
2002-01-01
期刊:
BMC pharmacology
影响因子:
--
作者:
[Kramer, H Kenneth, Onoprishvili, Irma, Simon, Eric J]
通讯作者:
Simon, Eric J
35th Annual International Narcotics Research Conference
-
批准号:6814513
-
项目类别:
-
资助金额:$5.6万
-
财政年份:2004
-
负责人:Eric J Simon
-
依托单位:
34th Annual International Narcotics Research Conference
-
批准号:6675986
-
项目类别:
-
资助金额:$5.25万
-
财政年份:2003
-
负责人:Eric J Simon
-
依托单位:
33rd International Narcotics Research Conference (INRC)
-
批准号:6531293
-
项目类别:
-
资助金额:$6.5万
-
财政年份:2002
-
负责人:Eric J Simon
-
依托单位:
32nd International Narcotics Research Conference(INRC)
-
批准号:6361861
-
项目类别:
-
资助金额:$6.5万
-
财政年份:2001
-
负责人:Eric J Simon
-
依托单位:
31ST ANNUAL INTERNATIONAL NARCOTICS RESEARCH CONFERENCE
-
批准号:6150456
-
项目类别:
-
资助金额:$5.0万
-
财政年份:2000
-
负责人:Eric J Simon
-
依托单位:
BIOCHEMICAL RESEARCH ON OPIOID RECEPTORS
-
批准号:2561669
-
项目类别:
-
资助金额:$10.59万
-
财政年份:1999
-
负责人:Eric J Simon
-
依托单位:
BIOCHEMICAL RESEARCH ON OPIOID RECEPTORS
-
批准号:6362781
-
项目类别:
-
资助金额:$12.71万
-
财政年份:1999
-
负责人:Eric J Simon
-
依托单位:
30TH ANNUAL INTERNATIONAL NARCOTICS RESEARCH CONFERENCE
-
批准号:2884096
-
项目类别:
-
资助金额:$6.0万
-
财政年份:1999
-
负责人:Eric J Simon
-
依托单位:
BIOCHEMICAL RESEARCH ON OPIOID RECEPTORS
-
批准号:6164331
-
项目类别:
-
资助金额:$12.71万
-
财政年份:1999
-
负责人:Eric J Simon
-
依托单位:
BIOCHEMICAL RESEARCH ON OPIOID RECEPTORS
-
批准号:6515292
-
项目类别:
-
资助金额:$12.71万
-
财政年份:1999
-
负责人:Eric J Simon
-
依托单位:
29TH ANNUAL INTERNATIONAL NARCOTIC RESEARCH CONFERENCE
-
批准号:2561677
-
项目类别:
-
资助金额:$6.56万
-
财政年份:1998
-
负责人:Eric J Simon
-
依托单位:
POSTDOCTORAL TRAINING IN RESEARCH ON ABUSED DRUGS
-
批准号:6378518
-
项目类别:
-
资助金额:$14.35万
-
财政年份:1991
-
负责人:Eric J Simon
-
依托单位:
POSTDOCTORAL TRAINING IN RESEARCH ON ABUSED DRUGS
-
批准号:2119619
-
项目类别:
-
资助金额:$11.74万
-
财政年份:1991
-
负责人:Eric J Simon
-
依托单位:
Postdoctoral Training in Research on Abused Drugs
-
批准号:7093058
-
项目类别:
-
资助金额:$21.13万
-
财政年份:1991
-
负责人:Eric J Simon
-
依托单位:
POSTDOCTORAL TRAINING IN RESEARCH ON ABUSED DRUGS
-
批准号:2119621
-
项目类别:
-
资助金额:$15.28万
-
财政年份:1991
-
负责人:Eric J Simon
-
依托单位:
POSTDOCTORAL TRAINING IN RESEARCH ON ABUSED DRUGS
-
批准号:2700853
-
项目类别:
-
资助金额:$15.86万
-
财政年份:1991
-
负责人:Eric J Simon
-
依托单位:
Postdoctoral Training in Research on Abused Drugs
-
批准号:6953677
-
项目类别:
-
资助金额:$21.44万
-
财政年份:1991
-
负责人:Eric J Simon
-
依托单位:
POSTDOCTORAL TRAINING IN RESEARCH ON ABUSED DRUGS
-
批准号:6174654
-
项目类别:
-
资助金额:$21.24万
-
财政年份:1991
-
负责人:Eric J Simon
-
依托单位:
POSTDOCTORAL TRAINING IN RESEARCH ON ABUSED DRUGS
-
批准号:2013075
-
项目类别:
-
资助金额:$15.95万
-
财政年份:1991
-
负责人:Eric J Simon
-
依托单位:
Postdoctoral Training in Research on Abused Drugs
-
批准号:6451022
-
项目类别:
-
资助金额:$26.4万
-
财政年份:1991
-
负责人:Eric J Simon
-
依托单位:
海外基金