The use of electrospray ionization MS to determine the structure of glycans in intact glycoproteins.
نویسندگان
چکیده
and VIIIb (I. M. Fearnley, unpublished work), and the cytochrome bc, subunit IX [16], have been recorded satisfactorily in conventional solvents containing acetonitrile or methanol, and in some cases in chloroform/methanol also [5]. However, it is not possible to record spectra of the large proteolipids such as ND2 and ATPase6 in solvents based on acetonitrile or methanol, as large hydrophobic proteins are insoluble in them. In the present work, it has been demonstrated that the chlorofordmethanol solvents are suitable for recording electrospray mass spectra of both small and large proteolipids, and, most importantly, that in spite of their hydrophobicity and low content of basic amino acids, large hydrophobic proteins can be analysed. These experiments open the way for measuring electrospray spectra of many membrane proteins that could not be analysed by this technique to date. An additional important advantage of the MS techniques, also demonstrated above, is that they have allowed hydrophobic proteins to be sequenced partially, by tandem MS procedures that avoid the difficulties associated with cleavage and purification of peptides from hydrophobic membrane proteins, as required by conventional sequencing methods. T h e power of these techniques is further illustrated by the analysis of a mixture of efrapeptins bound in a single crystal with F,-ATPase.
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ورودعنوان ژورنال:
- Biochemical Society transactions
دوره 24 3 شماره
صفحات -
تاریخ انتشار 1996