Biology Reference
In-Depth Information
5.
It is not mandatory to deglycosylate the cellular (SDS) extracts.
If deglycosylation is desired, it has to be done after protein
precipitation to remove SDS (see Subheading
3.5
).
6.
Because of the differential solubilization of extracellular pro-
teins, the 0.5 M NaCl and the 4 M guanidine extracts repre-
sent relatively simple subproteomes and can be effectively
analyzed by gel LC-MS/MS. Alternatively, they could be ana-
lyzed by DIGE, following the cleaning steps described above
(see Subheading
3.5
, step 1).
Acknowledgements
M.M. is a senior fellow of the British Heart Foundation.
References
1. Unlu, M., Morgan, M. E., and Minden, J. S.
(1997) Difference gel electrophoresis: a single
gel method for detecting changes in protein
extracts,
Electrophoresis 18
, 2071-2077.
2. Arrell, D. K., Neverova, I., and Van Eyk, J. E.
(2001) Cardiovascular proteomics: evolution
and potential,
Circ Res 88
, 763-773.
3. Didangelos, A., Simper, D., Monaco, C., and
Mayr, M. (2009) Proteomics of acute coronary
syndromes,
Curr Atheroscler Rep 11
, 188-195.
4. Mayr, M., Liem, D., Zhang, J., Li, X.,
Avliyakulov, N. K., Yang, J. I., Young, G.,
Vondriska, T. M., Ladroue, C., Madhu, B.,
Griffi ths, J. R., Gomes, A., Xu, Q., and Ping, P.
(2009) Proteomic and metabolomic analysis of
cardioprotection: Interplay between protein
kinase C epsilon and delta in regulating glucose
metabolism of murine hearts,
J Mol Cell Cardiol
46
, 268-277.
5. Mayr, M., Madhu, B., and Xu, Q. (2007)
Proteomics and metabolomics combined in
cardiovascular research,
Trends Cardiovasc Med
17
, 43-48.
6. Mayr, M., Zhang, J., Greene, A. S., Gutterman,
D., Perloff, J., and Ping, P. (2006) Proteomics-
based development of biomarkers in cardiovascu-
lar disease: mechanistic, clinical, and therapeutic
insights,
Mol Cell Proteomics 5
, 1853-1864.
7. McGregor, E., and Dunn, M. J. (2006)
Proteomics of the heart: unraveling disease,
Circ Res 98
, 309-321.
8. Yin, X., Cuello, F., Mayr, U., Hao, Z.,
Hornshaw, M., Ehler, E., Avkiran, M., and
Mayr, M. (2010) Proteomics analysis of the
cardiac myofi lament subproteome reveals
dynamic alterations in phosphatase subunit dis-
tribution,
Mol Cell Proteomics 9
, 497-509.
9. Didangelos, A., Yin, X., Mandal, K., Baumert,
M., Jahangiri, M., and Mayr, M. (2010)
Proteomics characterization of extracellular
space components in the human aorta,
Mol Cell
Proteomics 9
, 2048-2062.
10. Mason, R. M., and Mayes, R. W. (1973)
Extraction of cartilage protein-polysaccharides
with inorganic salt solutions,
Biochem J 131
,
535-540.
11. Dahl, S. L., Koh, J., Prabhakar, V., and Niklason,
L. E. (2003) Decellularized native and engi-
neered arterial scaffolds for transplantation,
Cell Transplant 12
, 659-666.
12. Korossis, S. A., Wilcox, H. E., Watterson, K.
G., Kearney, J. N., Ingham, E., and Fisher, J.
(2005) In-vitro assessment of the functional
performance of the decellularized intact por-
cine aortic root,
J Heart Valve Dis 14
, 408-421;
discussion 422.
13. Gilbert, T. W., Sellaro, T. L., and Badylak, S. F.
(2006) Decellularization of tissues and organs,
Biomaterials 27
, 3675-3683.
14. Sajdera, S. W., and Hascall, V. C. (1969)
Proteinpolysaccharide complex from bovine
nasal cartilage. A comparison of low and high
shear extraction procedures,
J Biol Chem 244
,
77-87.
15. Bannister, D. W., and Burns, A. B. (1972)
Pepsin treatment of avian skin collagen. Effects
on solubility, subunit composition and aggre-
gation properties,
Biochem J 129
, 677-681.
Search WWH ::
Custom Search