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. 2000 May 15;14(10):1236–1248.

Figure 2.

Figure 2

Molecular identification of p50 and p100. (A) Coomassie blue staining of purified p50 (lane 1) or p100 (lane 3) and their respective RNA–polypeptide complexes (lanes 2,4) detected by UV cross-linking and autoradiography. (B,C) Identification of YB-1 and nucleolin by the nanoelectrospray tandem mass spectra of doubly charged peptides extracted from the 50- and 100-kD bands, respectively. Sequence tags were assembled from a series of fragment ions and used to search for a matching pattern. Fragment masses calculated for the retrieved YB-1 GAEAANVTGPGGVPVQGSK or the retrieved nucleolin GYAFI/LEFASFEDAK sequences were compared with the complete fragmentation spectrum to confirm the match. (D) Schematic representation of nucleolin and YB-1. (Hatched boxes) RNA-binding domain; (dark grey box) RGG domain; (black boxes) RNP; (stippled boxes) cold shock domain.