Crystal structure of the FRP and identification of the active site for modulation of OCP-mediated photoprotection in cyanobacteria.

Photosynthetic reaction centers are sensitive to high light conditions, which can cause damage because of the formation of reactive oxygen species. To prevent high-light induced damage, cyanobacteria have developed photoprotective mechanisms. One involves a photoactive carotenoid protein that decreases the transfer of excess energy to the reaction centers.

Elucidating essential role of conserved carboxysomal protein CcmN reveals common feature of bacterial microcompartment assembly.

Bacterial microcompartments are organelles composed of a protein shell that surrounds functionally related proteins. Bioinformatic analysis of sequenced genomes indicates that homologs to shell protein genes are widespread among bacteria and suggests that the shell proteins are capable of encapsulating diverse enzymes. The carboxysome is a bacterial microcompartment that enhances CO(2) fixation in cyanobacteria and some chemoautotrophs by sequestering ribulose-1,5-bisphosphate carboxylase/oxygenase and carbonic anhydrase in the microcompartment shell.

Conformation-sensing antibodies stabilize the oxidized form of PTP1B and inhibit its phosphatase activity.

Protein tyrosine phosphatase 1B (PTP1B) plays important roles in downregulation of insulin and leptin signaling and is an established therapeutic target for diabetes and obesity. PTP1B is regulated by reactive oxygen species (ROS) produced in response to various stimuli, including insulin. The reversibly oxidized form of the enzyme (PTP1B-OX) is inactive and undergoes profound conformational changes at the active site.

Methods for preparing crystals of reversibly oxidized proteins: crystallization of protein tyrosine phosphatase 1B as an example.

Regulation of protein activity through the oxidation and reduction of cysteines is emerging as an important mechanism in the control of cell-signaling pathways. Protein tyrosine phosphatase 1B (PTP1B), for example, is reversibly inhibited by oxidation at the catalytic cysteine in response to stimulation of cells by insulin or epidermal growth factor. We have conducted structural studies on the redox regulation of PTP1B and have demonstrated that the oxidation of the catalytic cysteine results in the formation of a bond between the sulfur atom of the catalytic cysteine and the amide nitrogen of the neighboring serine.

Functions and mechanisms of redox regulation of cysteine-based phosphatases.

Reactive oxygen species (ROS) have been implicated as mediators of cell-signaling responses, particularly in pathways involving protein tyrosine phosphorylation. One mechanism by which ROS are thought to exert their effects is through the reversible regulation of cysteine-based phosphatases (CBPs). The CBPs, which include protein tyrosine phosphatases (PTPs), dual-specificity phosphatases, low-molecular-weight PTPs, and the lipid phosphatase PTEN, all contain a nucleophilic catalytic cysteine within a conserved motif that enables these enzymes to dephosphorylate phosphoproteins or phospholipids.

Redox regulation of protein tyrosine phosphatase 1B involves a sulphenyl-amide intermediate.

The second messenger hydrogen peroxide is required for optimal activation of numerous signal transduction pathways, particularly those mediated by protein tyrosine kinases. One mechanism by which hydrogen peroxide regulates cellular processes is the transient inhibition of protein tyrosine phosphatases through the reversible oxidization of their catalytic cysteine, which suppresses protein dephosphorylation. Here we describe a structural analysis of the redox-dependent regulation of protein tyrosine phosphatase 1B (PTP1B), which is reversibly inhibited by oxidation after cells are stimulated with insulin and epidermal growth factor.