Université de Fribourg

Chemical crosslinking and mass spectrometry to elucidate the topology of integral membrane proteins

Debelyy, Mykhaylo O. ; Waridel, Patrice ; Quadroni, Manfredo ; Schneiter, Roger ; Conzelmann, Andreas

In: PLOS ONE, 2017, vol. 12, no. 10, p. e0186840

Here we made an attempt to obtain partial structural information on the topology of multispan integral membrane proteins of yeast by isolating organellar membranes, removing peripheral membrane proteins at pH 11.5 and introducing chemical crosslinks between vicinal amino acids either using homo- or hetero-bifunctional crosslinkers. Proteins were digested with specific proteases and the...