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The next big thing in mass spectrometry
Chemistry World ^ | 8 March 2013 | David Bradley

Posted on 03/17/2013 2:58:54 PM PDT by neverdem

It's not quite the ‘elephant in the room’, but an 18 megadalton viral assembly is perhaps the biggest thing in the mass spectrometer (MS). Dutch and US researchers have used quadrupole time-of-flight (QToF) native MS to investigate intact capsids from a bacteriophage – a virus that infects bacteria. While there is theoretically no upper limit on the mass of a particle that might be analysed using ToF MS, the work is far from trivial in breaking through the record.

The late John Fenn shared the 2002 Nobel prize in chemistry for his pioneering work on electrospray ionisation techniques in mass spectrometry, which paved the way for it to handle larger and larger molecules. Critically, the techniques that have emerged during the last decade or so allow protein complexes and even viral particles to be carried unharmed into the gas phase where they can then be subjected to analysis. Native MS has recently been exploited in just such studies looking at intact viruses and viral capsids, which are important in the spread of viral disease but also as structures that might be exploited in nanotechnology and drug delivery.

The record-breaking virus capsid is made up of 420 40kDa sub-units, giving it an impressive molecular formula © Wiley-VCHAlbert Heck of Utrecht University in the Netherlands and colleagues there and at the Scripps Research Institute, La Jolla, California, have applied this analytical approach to bacteriophage HK97. This is an important model for understanding viruses that infect bacteria. They explain that the capsid assembles in vitro from a mixture of pentameric and hexameric capsomers made from protein gp5 and viral protease gp4. This builds an icosahedral intermediate, Prohead-1, which in nature matures by swelling up through activation of the protease molecules. However, the researchers could trap the growing capsid at the Prohead-1 stage for the investigation simply by leaving the proteases out of the recipe and over-expressing the gp5.

The results they have now obtained represent the biggest viral entity yet studied by mass spectrometry. The mass of the monomer building blocks are revealed to be above 40 kilodaltons, while the pentameric and hexameric capsomer assemblies are more than 210 kilodaltons and almost 253 kilodaltons, respectively, the team reports.

‘Given that the capsid comprises 420 gp5 units, we estimate the record-breaking mass to be close to 18 megadaltons. Indeed, we measured an experimentally accurate mass of 17.942+/-0.004 megadaltons,’ Heck tells Chemistry World.

The researchers suggest that there are various tweaks, such as improved desolvation, that might be made to a QToF instrument to further improve resolution allowing other protein complexes and viral particles as large as 20 MDa to be studied with good resolution.

Angela Corcelli, of the University of Bari Aldo Moro in Italy, is involved in complementary work that focuses on the lipid components of intact viruses. ‘Mass spectrometry of intact biological samples such as isolated membrane domains, organelles and viruses, represents an innovative analytical approach, which will provide novel extremely valuable information and data on biological structures,’ she says.

‘Heck's technique opens new exciting developments – not only in the study of proteins, but also in that of lipid-protein complexes and lipids of biomembranes.’

References

J Snijder et al, Angew. Chem., Int. Ed., 2013, DOI: 10.1002/anie.201210197



TOPICS: Science
KEYWORDS: bacteriophagehk97; massspectrometry; microbiology; virology

1 posted on 03/17/2013 2:58:54 PM PDT by neverdem
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To: neverdem

This sounds exciting and I’m happy but I didn’t understand a word of it.


2 posted on 03/17/2013 3:02:55 PM PDT by BipolarBob (Happy Hunger Games! May the odds be ever in your favor.)
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To: BipolarBob

Basically, they can detect that you farted without you knowing you have farted.


3 posted on 03/17/2013 3:11:30 PM PDT by Zeneta (No eternal reward will forgive us now for wasting the dawn.)
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To: neverdem

MASINT is ages old.


4 posted on 03/17/2013 3:13:26 PM PDT by mylife (The Roar Of The Masses Could Be Farts)
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To: Zeneta

Wow, I can see all kinds of uses for that information.


5 posted on 03/17/2013 3:15:48 PM PDT by BipolarBob (Happy Hunger Games! May the odds be ever in your favor.)
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To: neverdem

18 megadaltons is a lot of Daltons.


6 posted on 03/17/2013 3:31:47 PM PDT by rdl6989
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To: neverdem

Heap big molecule.


7 posted on 03/17/2013 3:34:28 PM PDT by Huskrrrr
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To: neverdem
I am certain you and many others found this extremely interesting but as for me, a simple man:


8 posted on 03/17/2013 4:27:27 PM PDT by A Formerly Proud Canadian (I once was lost but now I'm found; blind but now I see.)
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To: neverdem

Theoretically one could study a nano-analog of the entire universe using mass spectrometry.


9 posted on 03/17/2013 4:35:04 PM PDT by steve86 (Acerbic by Nature, not NurtureĀ™)
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To: BipolarBob

It can tell you which elements a material is made of.

Carbon, oxygen, hydrogen, etc. Each element has a “spectrum.”

Same idea when they look at stars and planets.

Apparently, they had troubles decoding more complicated molecules.


10 posted on 03/17/2013 4:37:05 PM PDT by dhs12345
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To: neverdem
Sounds interesting but the text is a little over my head. Is there a dumbed down version of this article?
11 posted on 03/17/2013 4:50:54 PM PDT by BenLurkin (This is not a statement of fact. It is either opinion or satire; or both)
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To: neverdem

bookmark.


12 posted on 03/17/2013 4:55:46 PM PDT by dadfly
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To: BenLurkin
Sounds interesting but the text is a little over my head. Is there a dumbed down version of this article?

Mass spectrometry allows not only detection but identification of molecules. The above work would allow relatively rapid id of viral materials.
13 posted on 03/17/2013 4:57:16 PM PDT by Nepeta
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To: Nepeta

I see. Thank you.


14 posted on 03/17/2013 5:05:46 PM PDT by BenLurkin (This is not a statement of fact. It is either opinion or satire; or both)
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To: dhs12345
"Apparently, they had troubles decoding more complicated molecules."

Not exactly. To "decode" a molecule (or other entity), you want the item to remain intact to the maximum extent possible (to get it's "parent mass"). The larger the molecules get, the more places they have available to break apart. If there is too much fragmentation, you lose the ability to extract the structural info.

Yet to get the molecule to work at all, you have to get it into the gas phase, remove at least one electron (to get it charged so the electric and magnetic fields can "steer" it, or accelerate it so that different mass fragments can be detected).

Past methods of ionization involved hitting the vaporizing molecules with a beam of electrons, or charged particles, which can be TOO energetic for the less stable giant molecules. So you need a kinder/gentler ionization means. This is one approach of several.

And the above is a VERY simplistic picture.

15 posted on 03/17/2013 5:15:40 PM PDT by Wonder Warthog
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To: rdl6989
No, this is a lot of Daltons...


16 posted on 03/17/2013 5:49:21 PM PDT by stormer
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To: neverdem

The viral particles can be carried unharmed into the gas phase? Thank goodness. We can send the SWAT team home.


17 posted on 03/17/2013 6:40:39 PM PDT by blueunicorn6 ("A crack shot and a good dancer")
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To: AdmSmith; AnonymousConservative; Berosus; bigheadfred; Bockscar; ColdOne; Convert from ECUSA; ...

Thanks neverdem.
...an 18 megadalton viral assembly is perhaps the biggest thing in the mass spectrometer (MS). Dutch and US researchers have used quadrupole time-of-flight (QToF) native MS to investigate intact capsids from a bacteriophage -- a virus that infects bacteria.

18 posted on 03/17/2013 7:16:23 PM PDT by SunkenCiv (Romney would have been worse, if you're a dumb ass.)
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To: stormer

If that doesn’t win you the post of the week award then I say the voting is rigged.


19 posted on 03/17/2013 7:29:43 PM PDT by rdl6989
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To: BipolarBob

identifying viral structures more readily may mean that they will find a way to kill the little buggers more easily like you know that viruses put up a protein shield to make them selves invisible to immune systems but i also read somewhere that there is work to make that shield visible to immune systems anyways the more they know about viruses the well and good we will all be in the longer run


20 posted on 03/17/2013 7:33:02 PM PDT by bigheadfred
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To: Zeneta

electrospray fartoid ionization mass spectrometry!


21 posted on 03/17/2013 8:06:41 PM PDT by Wanderer99
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To: neverdem

Boy, this is exciting stuff. Too bad it’s in a foreign language.


22 posted on 03/17/2013 8:15:57 PM PDT by Rocky (Obama is pure evil.)
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To: BipolarBob

Now, when some thug gets gunned down by an armed citizen defending himself from a mugging, the media can say, “He was an aspiring mass spectrometer.”


23 posted on 03/17/2013 8:21:40 PM PDT by Lancey Howard
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To: Wonder Warthog
Okay, but the molecule can’t be intact. At least in my limited understand of how a mass spec works you ionize the sample and then based on charge, separate the resulting ions.

How can you infer what the molecule is made of by preserving it? Without ionizing it?

Do you hit the molecule with different wavelength photons?

24 posted on 03/17/2013 8:27:18 PM PDT by dhs12345
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To: Rocky; dhs12345; BipolarBob; A Formerly Proud Canadian; BenLurkin
Rocky wrote, "Boy, this is exciting stuff. Too bad it’s in a foreign language."

Mass Spectrometry

One of the great things about the Internet is that you can look stuff up from reputable sources.

25 posted on 03/17/2013 9:42:36 PM PDT by neverdem ( Xin loi min oi)
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To: dhs12345
"Okay, but the molecule can’t be intact. At least in my limited understand of how a mass spec works you ionize the sample and then based on charge, separate the resulting ions."

"How can you infer what the molecule is made of by preserving it? Without ionizing it?

Correct. The problem is that TOO MUCH fragmentation loses needed information. You want at least SOME of the parent molecules to be intact or largely so, yet be ionized. So you have somewhat contradictory needs, and are working on balancing them.

And the bigger the molecule, the more difficult that balancing act gets.

"Do you hit the molecule with different wavelength photons?

AFAIK, that has not been used, or not widely used. Lasers HAVE been used to blast things off surfaces to get composition analysis, as with metal samples.

26 posted on 03/18/2013 5:10:54 AM PDT by Wonder Warthog
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To: Wonder Warthog

Cool. Thanks for explaining it to me.


27 posted on 03/19/2013 4:30:16 PM PDT by dhs12345
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To: neverdem

Cool. The closest I got to one of them was in Physics lab and of course, the theory behind it.


28 posted on 03/19/2013 4:34:23 PM PDT by dhs12345
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