Isolation of Peptides From Rat Tissues: Peptidomics vs. Degradomics
Earlier we have isolated about 200 functional protein fragments from acidic rat tissue extracts [1,2]. The conditions of their isolation (treatment time, t°C, absence of protease inhibitors) could indicate that at least part of those peptides resulted from post-mortem protein degradation. Such sets of protein fragments, or peptide pools, were shown to be tissue-specific and highly reproducible in individual animals their composition changing upon metabolic stress . Here we report isolation of peptides from rat brain, heart, lung and spleen extracts prepared in the presence of protease inhibitors and compare the resultant peptide sets with the earlier described sets of peptides.
Results and Discussion
The isolation of rat organs was performed within 1 min with immediate liquid nitrogen freezing. Tissues were homogenized in 10% acetic acid, in the presence or in absence of protease inhibitors (10−6M of pepstatin A, 10−4M of phenylmethylsulfonylfluoride (PMSF), 2 mM...
KeywordsPeptide Pool Packed Erythrocyte Nitrogen Freezing Edman Sequencing Liquid Nitrogen Freezing
The work was funded by RAS Presidium grant “Molecular and Cellular Biology”.