c**y
发帖数: 377 | 1 Can you give some comments on protein structure and property difference in
aqueous phase and gas phase?
thanks~~ | h******n
发帖数: 493 | 2 hehe, you probably want to talk to my boss about this.
He is an real expert but I am not. :)
There are still a lot of debates on whether the protein
structures and properties remain the same or change a
lot when the proteins go from agueous phase into gas
phase. The general idea is that structures and properties
of some proteins remain the same, but not all the proteins.
For some proteins, partial structures, especially the core
structures still remain unchanged. With the soft electrospray
ioniz
【在 c**y 的大作中提到】
: Can you give some comments on protein structure and property difference in
: aqueous phase and gas phase?
: thanks~~
| c**y
发帖数: 377 | 3 thanks, thanks. Reviews will be great, where are they?
Is there any different effect on non-covalent interactions, such as
electrostatic, H-bond, etc.? Do you have any comments on membrane associated
proteins?
【在 h******n 的大作中提到】
: hehe, you probably want to talk to my boss about this.
: He is an real expert but I am not. :)
: There are still a lot of debates on whether the protein
: structures and properties remain the same or change a
: lot when the proteins go from agueous phase into gas
: phase. The general idea is that structures and properties
: of some proteins remain the same, but not all the proteins.
: For some proteins, partial structures, especially the core
: structures still remain unchanged. With the soft electrospray
: ioniz
| h******n
发帖数: 493 | 4 There are certainly different effects on different types
of interactions. Electrostatic interactions are usually
strengthen in the gas phase, while the effects on H-bond
and hydrophobic interactions are not clear. I can't really
say too much about this, since I am not knowledged enough.:)
References:
1) Loo, J. A. (1997)
Studying noncovalent protein complexes by
electrospray ionization mass spectrometry.
Mass Spectrom Rev 16, 1-23
2) Loo, J. A. (2000)
Electrospray ionization mass spectrometry: a
【在 c**y 的大作中提到】
: thanks, thanks. Reviews will be great, where are they?
: Is there any different effect on non-covalent interactions, such as
: electrostatic, H-bond, etc.? Do you have any comments on membrane associated
: proteins?
| c**y
发帖数: 377 | 5 thanks again!
associated
【在 h******n 的大作中提到】
: There are certainly different effects on different types
: of interactions. Electrostatic interactions are usually
: strengthen in the gas phase, while the effects on H-bond
: and hydrophobic interactions are not clear. I can't really
: say too much about this, since I am not knowledged enough.:)
: References:
: 1) Loo, J. A. (1997)
: Studying noncovalent protein complexes by
: electrospray ionization mass spectrometry.
: Mass Spectrom Rev 16, 1-23
| m*******h
发帖数: 896 | 6 Man, you need also list some other references ah ;)
【在 h******n 的大作中提到】
: There are certainly different effects on different types
: of interactions. Electrostatic interactions are usually
: strengthen in the gas phase, while the effects on H-bond
: and hydrophobic interactions are not clear. I can't really
: say too much about this, since I am not knowledged enough.:)
: References:
: 1) Loo, J. A. (1997)
: Studying noncovalent protein complexes by
: electrospray ionization mass spectrometry.
: Mass Spectrom Rev 16, 1-23
| c**y
发帖数: 377 | 7 Is there any reference supporting or objecting the idea of using MS for
analyzing protein-protein interaction? for example, you have protein A, and
after you overexpress A in a common cell line, you use antibody
for A to fish any protein that could bind A, and use Mass to detect, how
efficient you can get? Is the result trustable?
by the way, i am not doing anything with this, just curious.
associated
【在 m*******h 的大作中提到】
: Man, you need also list some other references ah ;)
| h******n
发帖数: 493 | 8 It is useless to list too many. Two is enough. ;)
【在 m*******h 的大作中提到】
: Man, you need also list some other references ah ;)
| h******n
发帖数: 493 | 9 This is a very common method and called immunoprecipitation.
【在 c**y 的大作中提到】
: Is there any reference supporting or objecting the idea of using MS for
: analyzing protein-protein interaction? for example, you have protein A, and
: after you overexpress A in a common cell line, you use antibody
: for A to fish any protein that could bind A, and use Mass to detect, how
: efficient you can get? Is the result trustable?
: by the way, i am not doing anything with this, just curious.
:
: associated
| c**y
发帖数: 377 | 10 Maybe i didn't make the question clear, what i am asking is whether Mass is
efficient to detect the possible interaction? Or, how the results from Mass
corelate with physiological function?
and
【在 h******n 的大作中提到】
: This is a very common method and called immunoprecipitation.
| h******n
发帖数: 493 | 11 The sensitivity of detecting the intact protein
complexes by MS directly is still not very good,
not as good as detecing just proteins under the
denatured conditions.
But people usually immunoprecipitate the protein
complexes and then run gels or use other methods
to separate the proteins, and then identify the
ligand proteins by MS. This method is very sensitive
and accurate.
Hope this answer your question.
【在 c**y 的大作中提到】
: Maybe i didn't make the question clear, what i am asking is whether Mass is
: efficient to detect the possible interaction? Or, how the results from Mass
: corelate with physiological function?
:
: and
| c**y
发帖数: 377 | 12 thanks~ That helps
is
Mass
【在 h******n 的大作中提到】
: The sensitivity of detecting the intact protein
: complexes by MS directly is still not very good,
: not as good as detecing just proteins under the
: denatured conditions.
: But people usually immunoprecipitate the protein
: complexes and then run gels or use other methods
: to separate the proteins, and then identify the
: ligand proteins by MS. This method is very sensitive
: and accurate.
: Hope this answer your question.
| c******n
发帖数: 16403 | 13 在厦大的时候大多都学物化, 出来后好像学物化的比学分析有机的都要少.
【在 c**y 的大作中提到】
: Can you give some comments on protein structure and property difference in
: aqueous phase and gas phase?
: thanks~~
| m*******h
发帖数: 896 | 14 I am doing something like this.
The problem in this kind of study is not the efficiency of MS, for MS is
sensitive enough for subfemtomole detection.
The major problem in this study is the "Non-specific" binding.
Your back groud will be pretty high, and you basically have no way
to distinguish the "back ground" and the "specific interaction"
In this year, John Yates published a mini review about "Complexomes
Studies by MS" on Molecular & Cellular Proteomics, a new journal for
proteomics study. Y
【在 c**y 的大作中提到】
: Is there any reference supporting or objecting the idea of using MS for
: analyzing protein-protein interaction? for example, you have protein A, and
: after you overexpress A in a common cell line, you use antibody
: for A to fish any protein that could bind A, and use Mass to detect, how
: efficient you can get? Is the result trustable?
: by the way, i am not doing anything with this, just curious.
:
: associated
|
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