Determination of protein-protein interactions through aldehyde-dextran intermolecular cross-linking.
A very simple strategy, based on the intermolecular cross-linking of associated proteins by using aldehyde-dextrans, has been proposed to detect protein-protein interactions. Aldehyde-dextran was able to cross-link different enzymes composed of several polypeptide chains (e.g., trypsin and penicillin G acylase), proteolyzated proteins (e.g., extracts from porcine pancreas) and finally, an immunocomplex (horseradish peroxidase/anti-horseradish peroxidase). This cross-linked immunocomplex could be selectively adsorbed on immobilized anti-rabbit IgG. The presence of unspecific covalent attachment between unrelated protein molecules was not detected. Thus, this strategy permits the cross-linking of different protein components and avoids the formation of nonspecific protein-protein associations.
Fuentes
Manuel
M
Departamento of Biocatálisis, Instituto de Catálisis (CSIC), Campus UAM Cantoblanco, Madrid, Spain.
Segura
Rosa L
RL
Abian
Olga
O
Betancor
Lorena
L
Hidalgo
Aurelio
A
Mateo
Cesar
C
Fernandez-Lafuente
Roberto
R
Guisan
Jose M
JM
D000447:Aldehydes / Q000737:chemistry*; D000818:Animals; D003432:Cross-Linking Reagents / Q000737:chemistry*; D003911:Dextrans / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D008958:Models, Molecular; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D011817:Rabbits
D000447:Aldehydes; D003432:Cross-Linking Reagents; D003911:Dextrans; D011506:Proteins