Strategies for the purification of membrane proteins.

نویسنده

  • Sinead Marian Smith
چکیده

Although membrane proteins account for 20-30% of the coding regions of all sequenced genomes and play crucial roles in many fundamental cell processes, there are relatively few membranes proteins with known 3D structure. This is likely due to technical challenges associated with membrane protein extraction, solubilisation, and purification. Membrane proteins are classified based on the level of interaction with membrane lipid bilayers, with peripheral membrane proteins associating non-covalently with the membrane, and integral membrane proteins associating more strongly by means of hydrophobic interactions. Generally speaking, peripheral membrane proteins can be purified by milder techniques than integral membrane proteins, whose extraction requires phospholipid bilayer disruption by detergents. Here, important criteria for strategies of membrane protein purification are addressed, with a focus on the initial stages of membrane protein solublilisation, where problems are most frequently encountered. Protocols are outlined for the successful extraction of peripheral membrane proteins, solubilisation of integral membrane proteins, and detergent removal which is important not only for retaining native protein stability and biological functions, but also for the efficiency of later purification techniques.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Practical Techniques for Improving the Performance of Polymeric Membranes and Processes for Protein Separation and Purification

Protein separation and purification technologies play an essential role in various industries including but not limited to pharmaceuticals, dairy as well as the food sector. Accordingly, a wide variety of techniques such as chromatography and electrophoresis has been developed and utilized extensively over the years for this purpose. Despite their widespread acceptance, conventiona...

متن کامل

ISOLATION AND PURIFICATION OF MAJOR OUTER MEMBRANE PROTEINS FROM BRUCELLA ABORTUS S-99

Isolation and purification of major outer membrane proteins (OMP) from the cell wall envelope of Brucella abortus S-99 were achieved by sonication, solubilization and membrane fractionation in the presence of non-ionic detergent (Tx-100) and lysozyme treatments, followed by ultracentrifugation. The crude OMP was treated with trypsin to free the preparation from any other protein contaminan...

متن کامل

Affinity Based Nano-Magnetic Particles for Purification of Recombinant Proteins in Form of Inclusion Body

Background: Protein purification is the most complicated issue in the downstream processes of recombinant protein production; therefore, improved selective purification methods are important. Affinity-based protein purification method using His-tag and Ni-NTA resins is one of the most common strategies. MNPs can be used as a beneficial alternative for Ni-NTA resins. However, there is no data on...

متن کامل

Evaluation and comparison of affinity chromatography and precipitation– based methods on purification of recombinant streptokinase

Background: Increase of protein purity is a serious challenge in the production of recombinant therapeutic proteins. For this purpose, several strategies have been employed to purify the target protein, among which the affinity chromatography-based purification methods and tagged proteins such as Ni-NTA are common and but costly. Therefore column-free purification techniques, such as using elas...

متن کامل

Evaluation of Porin Interaction with Adenine Nucleotide Translocase and Cyclophilin-D Proteins after Brain Ischemia and Reperfusion

Objective (s) Porin is a mitochondrial outer membrane channel, which usually functions as the pathway for the movement of various substances in and out of the mitochondria and is considered to be a component of the permeability transition (PT) pore complex that plays a role in the PT. We addressed the hypothesis that porin interacts with other mitochondrial proteins after ischemic injury. Mater...

متن کامل

Cloning and expression of Brucella outer membrane protein 36kDa (OMP2b) in E. coli

Background & Objective: Brucellosis is an important zoonotic disease of economic significance. Brucella species are gram-negative, facultative intracellular bacteria, and are capable of replicating in the phagosomes of macrophages. They cause infection in several animal species and humans. Prevention of new diseases and diagnosis of cases infected with the organism are both essential for eradic...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Methods in molecular biology

دوره 1485  شماره 

صفحات  -

تاریخ انتشار 2011