PHYSICAL AND GENETIC STUDIES OF REPRESSOR PROTEINS
PHYSICAL AND GENETIC STUDIES OF REPRESSOR PROTEINS
批准号:
3271142
负责人:
KATHLEEN S MATTHEWS
金额:
$15.02万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
1979
资助国家:
美国
项目状态:
已结题
起止时间:
1979-04-01 至 1992-03-31
关键词:
DNA Escherichia coli X ray crystallography analytical method antibody bacterial genetics calorimetry chemical association chemical reaction chemical structure function circular DNA conformation fluorescence spectrometry lac operon mutant nuclear magnetic resonance spectroscopy point mutation thermodynamics tryptophan
中文摘要
这个研究项目的目的是调查机制
英文摘要
This research project is designed to investigate the mechanisms by which
proteins recognize specific sequences of DNA and the means by which small
ligands modulate the recognition process. The lac and trp repressors from
E. coli represent regulatory proteins in the classes of inducible and
repressible systems; both of these proteins and their target DNA sequences
can be isolated in the quantities required for physical and chemical
studies, and the coding regions are available on vectors for site-specific
mutagenesis and other genetic manipulations. The specific objectives are:
(1) assessment of the roles of specific amino acids and regions of the
primary sequences in the function of these repressor proteins. This
examination will include generation of site-specific mutants based on the
trp repressor x-ray crystallographic structure and on the known chemical,
physical and homology data for lac repressor. Interchange of the
helix-turn-helix structure implicated in DNA binding between the two
repressors will be undertaken. These specifically designed proteins and
mutant proteins available from other sources will be characterized in
detail. Antibodies to specific segments of the primary sequence of these
proteins will be generated, and the effects of antibody-repressor complex
formation on functional activities will be measured. (2) Determination of
the effects of DNA structure on binding activities. The effects of
supercoiling on the binding parameters for repressor-DNA (both operator and
nonspecific binding) will be examined. Cross-linking of proteins to DNA
will be attempted to assess specific regions of the protein which are in
close proximity to DNA. (3) Thermodynamic studies of protein structure and
function. High pressure dissociation of the repressor proteins and effects
of ligands on this process will provide information about the strength of
inter-subunit contacts and alterations in these contacts by ligand
binding. Calorimetric measurements of protein denaturation (DSC) and
ligand binding will be undertaken to assess the thermodynamic properties of
the proteins and their interactions. (4) Determination of protein
structure and conformational changes. FLuorescence spectroscopy of
intrinsic and extrinsic fluorophores will be measured, and NMR spectroscopy
will be utilized to examine the binding properties of the trp repressor.
The correlation of data obtained from these studies on two different
systems with data available in the literature will expand our knowledge of
genetic regulation, an essential function in all living organisms.
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Allosteric Transition in Lactose Repressor Protein
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批准号:7928481
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项目类别:
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资助金额:$4.61万
-
财政年份:2009
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
From genetic architecture to adaptation dynamics
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批准号:7267714
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项目类别:
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资助金额:$28.33万
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财政年份:2004
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负责人:KATHLEEN S MATTHEWS
-
依托单位:
From genetic architecture to adaptation dynamics
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批准号:7478548
-
项目类别:
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资助金额:$28.33万
-
财政年份:2004
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
SMALL INSTRUMENTATION GRANT
-
批准号:3525770
-
项目类别:
-
资助金额:$3.29万
-
财政年份:1992
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
SMALL INSTRUMENTATION GRANT
-
批准号:3525702
-
项目类别:
-
资助金额:$1.59万
-
财政年份:1991
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
SMALL INSTRUMENTATION GRANT
-
批准号:3523074
-
项目类别:
-
资助金额:$1.86万
-
财政年份:1990
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
SMALL INSTRUMENTATION PROGRAM
-
批准号:3524762
-
项目类别:
-
资助金额:$1.96万
-
财政年份:1989
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
PURCHASE OF FERMENTER AND OLIGONUCLEOTIDE SYNTHESIZER
-
批准号:3519312
-
项目类别:
-
资助金额:$22.0万
-
财政年份:1986
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
MUTANTS OF REPRESSOR AND PERIPLASMIC BINDING PROTEINS
-
批准号:3287296
-
项目类别:
-
资助金额:$14.61万
-
财政年份:1985
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
MUTANTS OF REPRESSOR AND PERIPLASMIC BINDING PROTEINS
-
批准号:3287297
-
项目类别:
-
资助金额:$17.03万
-
财政年份:1985
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
MUTANTS OF REPRESSOR AND PERIPLASMIC BINDING PROTEINS
-
批准号:3287298
-
项目类别:
-
资助金额:$12.11万
-
财政年份:1985
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
MOLECULAR AND CELL BIOCHEMISTRY AND BIOLOGY
-
批准号:3537963
-
项目类别:
-
资助金额:$6.88万
-
财政年份:1980
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
TRAINING PROGRAM IN MOLECULAR & CELL BIOCHEMISTRY & BIOL
-
批准号:2166967
-
项目类别:
-
资助金额:$8.75万
-
财政年份:1980
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
MOLECULAR AND CELL BIOCHEMISTRY AND BIOLOGY
-
批准号:3537964
-
项目类别:
-
资助金额:$6.88万
-
财政年份:1980
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
TRAINING PROGRAM IN MOLECULAR & CELL BIOCHEMISTRY & BIOL
-
批准号:2166968
-
项目类别:
-
资助金额:$8.96万
-
财政年份:1980
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
PHYSICAL AND GENETIC STUDIES OF REGULATORY PROTEINS
-
批准号:2173933
-
项目类别:
-
资助金额:$30.15万
-
财政年份:1979
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
PHYSICAL AND GENETICS STUDIES OF REGULATORY PROTEINS
-
批准号:2684680
-
项目类别:
-
资助金额:$30.32万
-
财政年份:1979
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
PHYSICAL AND GENETIC STUDIES OF REPRESSOR PROTEINS
-
批准号:3271147
-
项目类别:
-
资助金额:$15.2万
-
财政年份:1979
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
PHYSICAL AND GENETIC STUDIES OF REGULATORY PROTEINS
-
批准号:3271143
-
项目类别:
-
资助金额:$27.36万
-
财政年份:1979
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
BIOMEDICAL RESEARCH SUPPORT
-
批准号:3518365
-
项目类别:
-
资助金额:$6.19万
-
财政年份:1979
-
负责人:KATHLEEN S MATTHEWS
-
依托单位:
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