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| Institute: |
Lab or Branch |
| Waters Corporation, Life Sciences R&D
(CRD), Milford MA |
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| Title: |
| Dramatically enhanced proteolytic digestion
of proteins employing a novel denaturant |
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| Authors: |
| J.C. Gebler, Y-Y. Yu, M. Gilar, and P.
J. Lee |
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| Abstract: |
| The novel surfactant (sodium 4-[2-methyl-2-undecyl-1,3-dioxolan-4-yl)
methoxyl]-1-propane sulfonate (trade name RapiGest™)
has been found to be very useful as an aid during the
proteolytic digestion of proteins. In addition, the reagent
is acid-labile making its removal simple. The denaturant
has been found especially useful in accelerating digestion
of proteins in the presence of trypsin. For example, the
globular protein Myoglobin can be completely digested
in less then 20 minutes with 0.05% RapiGest™ (control
was 20 hours). Since the reagent can be removed it does
not interfere with post analysis methods such as HPLC,
MALDI-MS, and LC/MS. Approaches will be presented for
the complete enzymatic digestion of very hydrophobic proteins
and proteolytic resistant proteins. Data will be shown
where the surfactant was employed to facilitate the complete
tryptic digest of the hydrophobic protein Bacteriophodopsin.
Compatibility of the denaturant with other common proteases,
Asp-N, Glu-C, Lys-C, and chymotrypsin will be presented. |
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