On 2014 Dec 01, Cosmin Saveanu commented:

Affinity purification of pre-60S complexes associated with tagged Ssf1, Rix1, Arx1 and Lsg1 (alias Kre35) coupled with quantitative mass spectrometry were used to obtain a picture of pre-60S dynamics in yeast. Overexpression of a dominant negative form of Drg1, as well as deletion of BUD20 were also used to perturb 60S assembly and test the changes in levels of various pre-60S factors.

The quantitations presented in this paper correlate well with our published results that were based on affinity purification of Rlp24, Nog1 and Mak11 associated particles under various mutant conditions and quantitative mass-spectrometry based on stable isotope labeling (Lebreton A, 2008).

The major caveat of these methods is the uncertainity in the faith of blocked pre-60S particle that accumulate under mutant conditions - are these "dead-end" or real intermediates in the pathway ?

On 2014 Dec 01, Cosmin Saveanu commented:

Affinity purification of pre-60S complexes associated with tagged Ssf1, Rix1, Arx1 and Lsg1 (alias Kre35) coupled with quantitative mass spectrometry were used to obtain a picture of pre-60S dynamics in yeast. Overexpression of a dominant negative form of Drg1, as well as deletion of BUD20 were also used to perturb 60S assembly and test the changes in levels of various pre-60S factors.

The quantitations presented in this paper correlate well with our published results that were based on affinity purification of Rlp24, Nog1 and Mak11 associated particles under various mutant conditions and quantitative mass-spectrometry based on stable isotope labeling (Lebreton A, 2008).

The major caveat of these methods is the uncertainity in the faith of blocked pre-60S particle that accumulate under mutant conditions - are these "dead-end" or real intermediates in the pathway ?