Bifunktionelle Reagentien zur Quervernetzung von Proteinen

Abstract

Proteine konnen durch chemische Verbindungen quervernetzt werden, ohne daJ die komplizierte Raumstruktur sich dabei iindert. Man kennt inzwischen uiele bifunktionelle Reagentien, die mit funktionellen Gruppen in den Seitenketten der Aminosauren zu reagieren vermogen. Aus der bekannten Lange der eingefihrten Briicke kann nicht nur auf den Abstand der verkniipfen Aminosaurereste geschlossen werden, sondern es lassen sich u. a. auch Aussagen iiber Konformationsanderungen wahrend anderer Reaktionen und iiber die Anordnung der Bestandteile einer Quartarstruktur gewinnen.

Analysis of the reconstitution of oligomeric enzymes by crosslinking with glutaraldehyde: kinetics of reassociation of lactic dehydrogenase

“…Chemical cross-linking has become a ubiquitous tool in protein chemistry, its application including protein topography (Fasold et al, 1971; Wold, 1972), conformational transitions (Enns & Chan, 1978;Klemes & Citri, 1980), stoichiometry of subunits in oligomeric proteins (Davies & Stark, 1970) and their spatial arrangement (Bickle et al, 1972;Hucho et al, 1975;Hajdu et al, 1976;Friedrich et al, 1979), and fixation of proteins in the crystalline state (Quiocho & Richards, 1964), among others. In a previous paper, chemical cross-linking by glutaraldehyde was introduced as a new approach in studying the association kinetics of oligomeric systems (Hermann et al, 1979).…”

A simplified route to [14C] dimethylsuberimidate for polypeptide crosslinking studies

“…Crosslinking reagents have become valuable tools in the study of the interactions between subunits in multimeric proteins [ I], in determining the proximity of segments of the same polypeptide chain [2] or, as in the study of prokaryotic ribosomes, the interactions between contiguous proteins [3]. The analytical potential of these reagents in assessing the symmetry of tetrameric enzymes, for which X-ray diffraction data may not be available, has already been demonstrated [4].…”

5‐Isothiocyanato‐1‐naphthalene Azide and p‐Azidophenylisothiocyanate

“…The spatial arrangement of complex biological structures [I -31 and the molecular folding of proteins [4] have been investigated by cross-linking reagents varying in size, specificity and topological selectivity. Bifunctional cross-linking reagents which specifically bind to membrane proteins in the first covalent step prior to the more unspecific reaction of the hetero function will produce data which are easier to interpret.…”