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Specifically, the developments comprise (1) higher\u2010sensitivity chemical sequencing through background reduction; (2) improved peptide recovery from rapid in situ digests of nanogram amount, nitrocellulose\u2010bound proteins; and (3) accurate UV spectroscopic identification of Trp\u2010 and Cys\u2010containing peptides. In addition, we describe strategies for 2\u2010dimensional liquid chromatographic peptide isolation from complex mixtures and a multi\u2010analytical approach to peptide sequence analysis (Edman sequencing, matrix\u2010assisted laser desorption mass spectrometry, and UV spectroscopy). Both strategies were applied in tandem to the primary structural analysis of a gel\u2010purified, 250\u2010kDa protein (mammalian target of rapamycin\u2010FKBP12 complex), available in low picomolar quantities only. More than 300\u2010amino acids worth of sequence was obtained in mostly uninterrupted stretches, several containing Trp, Cys, His, and Ser. That information has allowed the matching of a biological function of a mammalian protein to a yeast gene product with a well\u2010characterized mutant phenotype. The results also demonstrate that extended chemical sequencing analysis (e.g., 26 successive amino acids) is now feasible, starting with initial yields well below 1 pmol.<\/jats:p>","DOI":"10.1002\/pro.5560031227","type":"journal-article","created":{"date-parts":[[2009,2,10]],"date-time":"2009-02-10T01:29:19Z","timestamp":1234229359000},"page":"2435-2446","source":"Crossref","is-referenced-by-count":44,"title":["High\u2010Sensitivity sequencing of large proteins: Partial structure of the rapamycin\u2010fkbp12 target"],"prefix":"10.1002","volume":"3","author":[{"given":"Hediye","family":"Erdjument\u2010Bromage","sequence":"first","affiliation":[]},{"given":"Mary","family":"Lui","sequence":"additional","affiliation":[]},{"given":"Paul","family":"Tempst","sequence":"additional","affiliation":[]},{"given":"David M.","family":"Sabatini","sequence":"additional","affiliation":[]},{"given":"Solomon 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