A fa Anal is very different from Protein interactions yzing proteins Is carried out indirectly by the so-called pull-down assays. To do this, the immobilized protein of interest to an affinity Tss molecules AT7519 To anything similar manner to the methods of affinity Create tschromatographie. After all bound proteins Were captured and released MS analysis is h Frequently after separation by gel electrophoresis and digestion 1D separated proteins Performed. Instead, affinity purification Tschromatographie based complex normal protein Immunpr Zipitation or affinity tsreinigung Used in tandem. Discussed proteins Fishing MS-based Ans tze In this article is not suitable for the study of dynamic protein binding events, but t would allow the identification of important multi-protein complexes with many different proteins.
After all, the SPR was with Member States coupled protein binding events on a chip SPR directly followed by MS identification of related proteins to investigate. This approach makes Glicht us to quantify the protein having the structural characterization / identification of proteins. As a result, the full MS and SPR detection show structural Ver Changes not detected by SPR. The study of non-covalent complexes of MS analysis of non-covalent complexes of native MS, also called native MS is known, requires ESI buffers compatible. This implies that in a number of F Cases the maximum sensitivity is not achieved and / or to use non-physiological conditions. Although not non-covalent complexes are observed by the Member States directly from real cellular Other systems, mostly the St Stoichiometry of the complexes by MS native games analyzed by other means, such as electron microscopy was determined, R ntgenkristallographie And NMR.
It is recognized, however, some exceptions. In general, native MS is a very POWERFUL Hige technique to study protein complexes, complement R Ans tze more traditional. Other Ans tze Like crystallography and NMR analysis functions. W While crystallography provides a detailed 3D image of a protein-ligand complex is obtained, the analysis of multi-protein complexes, complexes, and several types of classes of proteins in general difficult or protein complexes m Possibly the unm Crystallize resembled . In addition, the crystallographic analysis of the crystal structures of both static and dynamic analysis of the actual product is not chlichen in vitro m Possible.
Protein NMR, on the other hand, is a new technology, but the study of protein structure large and complex assemblies such as e whole virus has not m Is possible. NMR of proteins, proteins were In size Investigated enbereich of 0.1 MDa first With NMR studies homomeric complexes are lighter than heteromeric protein complexes analyzed NMR spectra become much more difficult to interpret, that is not a problem with native MS. Currently one of the largest are Th protein structure by NMR transcarbamylase proteins Gel 300 kDa and 670 kDa st aspartate proteasome by the group of Kay received. With native MS is one of the gr Investigated th assembly of protein complexes currently Norwalk virus. All technologies used to study protein complexes have their own advantages and disadvantages.