Analysis of Protein Localization in Plant Endoplasmic Reticulum

In plant cells, the secretory system delivers many different proteins to four major locations, including the cell wall, vacuole, plasma membrane, and tonoplast. The newly synthesized protein must have a specific conformation in the ER before further downstream delivery by the secretory system. Peptides that fail to fold correctly or never become part of the multi-subunit complex are retained in this compartment. The cells perform 'quality control' on the newly synthesized peptides, allowing the elimination of non-functional proteins.

Lifeasible develops an advanced platform that provides services to customers worldwide covering analysis of protein localization in plant endoplasmic reticulum. We customize featured services according to customer needs with decades of experience in plants. In addition, we deliver satisfactory and reliable results and reports on time to our customers worldwide.

Signal-Independent Retention in ER

• For a small number of resident proteins, their retention is independent of the signal.
• It is reported that proteins with specific ER-retention sequences reside in the ER through prolonged interactions with molecular chaperones such as BiP.
• In practice, we achieve this goal via establishing of protein mutants, the transformation of plants, production of the anti-Bip antiserum, total leaf protein extraction, subcellular fractionation, protein gel blot analysis, protoplast preparation, pulse-chase labeling, immunoprecipitation, the transient transformation of protoplasts, quantification of radioactive protein and so on. We always propose solutions according to the specific situation of your experiment.

Localization Signals for Soluble, ER-Resident Proteins

• In plants, both HDEL and KDEL C-terminal extensions have been identified in the sequences of soluble, ER-resident proteins that are collectively referred to as reticuloplasmins. In addition, it is reported that the addition of KDEL to the C-terminal end of various secreted proteins leads to the retention of these in the ER.

Fig.1 Possible mechanisms of ER protein retention.

• Based on the above principle, we provide services to illustrate the point, including chimeric the target gene, plant transformation, protein extraction, endoglycosidase-H digestion, immunoblotting, and immunocytochemistry.

Retention of Integral Membrane ER Proteins

• Although the default route for the vectorial flow of membrane proteins is from the ER to the plasma membrane or vacuole membrane, specific signals and mechanisms are necessary to retain resident proteins within the ER or Golgi membrane.
• Based on the above theory, we provide services to prove this view, including complementary cloning of plant ER type I membrane proteins and introduction into plant of both wild-type and mutant plants by Ti plasmid-mediated transformation.

Lifeasible offers professional services covering analysis of protein localization in the plant ER to meet your research needs. With years of experience in plants, our advanced platforms can help our clients solve various difficulties and conduct research. If you are interested in our services or have any questions, please feel free to contact us or make an online inquiry.