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Title (Primary) Determination of disulfide bond patterns in laminin β1 chain N-terminal domains by nano-high-performance liquid chromatography/matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry
Author Kalkhof, S.; Haehn, S.; Ihling, C.; Paulsson, M.; Smyth, N.; Sinz, A.;
Journal Rapid Communications in Mass Spectrometry
Year 2008
Department PROTEOM;
Volume 22
Issue 12
Language englisch;
Abstract The disulfide bonding patterns in the N-terminal (LN) domains of the basement membrane protein laminin beta 1 have not been investigated so far. We report an in-depth mass spectrometric analysis using offline nano-high-performance liquid chromatography/matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry (nano-HPLC/MALDI-TOF/TOF-MS) for determining the disulfide bond patterns in the LN-domain of recombinant mouse laminin beta 1 chain for the first time. Mass spectra were recorded and the putatively disulfide-linked peptides were subjected to LIFT-TOF/TOF-MS to confirm the disulfide bond. Screening the fragment ion mass spectra of disulfide-linked peptides for characteristic 66-amu patterns (34 u +32 u), arising from symmetric and asymmetric cleavage of disulfide bonds, facilitated their identification. Using various enzymes for proteolytic digestion of a recombinant laminin beta 1 chain N-terminal protein fragment, a linear bonding pattern of the eight cysteine residues in the LN-domain of the laminin beta 1 chain was observed with a (1-2, 3-4, 5-6, 7-8) connectivity of cysteines. The identical disulfide-bonding pattern was found in E4, the N-terminal laminin beta 1 chain fragment derived by elastase digestion of mouse tumor laminin-111, confirming that this pattern also occurs in native laminin. Copyright (C) 2008 John Wiley & Sons, Ltd
ID 1084
Persistent UFZ Identifier https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=1084
Kalkhof, S., Haehn, S., Ihling, C., Paulsson, M., Smyth, N., Sinz, A. (2008):
Determination of disulfide bond patterns in laminin β1 chain N-terminal domains by nano-high-performance liquid chromatography/matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry
Rapid Commun. Mass Spectrom. 22 (12), 1933 - 1940