Detection and quantification of free sulfhydryls in monoclonal antibodies using maleimide labeling and mass spectrometry.
AuthorsRobotham AC, Kelly JF
Journal
PubMed ID30894096
The detection of free sulfhydryls in proteins can reveal incomplete disulfide bond formation, indicate cysteine residues available for conjugation, and offer insights into protein stability and structure. Traditional spectroscopic methods of free sulfhydryl detection, such as Ellman's reagent, generally require a relatively large amount of sample, preventing their use for ... More
Regulating G protein-coupled receptors by topological inversion.
AuthorsDenard B, Han S, Kim J, Ross EM, Ye J
JournalElife
PubMed ID30835201
'G protein-coupled receptors (GPCRs) are a family of proteins containing seven transmembrane helices, with the N- and C-terminus of the protein located at the extracellular space and cytosol, respectively. Here, we report that ceramide or related sphingolipids might invert the topology of many GPCRs that contain a GXXXN motif in ... More
Transducin activates cGMP phosphodiesterase by trapping inhibitory ? subunit freed reversibly from the catalytic subunit in solution.
AuthorsAsano T, Kawamura S, Tachibanaki S
JournalSci Rep
PubMed ID31076603
'Activation of cGMP phosphodiesterase (PDE) by activated transducin a subunit (Ta*) is a necessary step to generate a light response in vertebrate photoreceptors. PDE in rods is a heterotetramer composed of two catalytic subunits, PDEa and PDEß, and two inhibitory PDE? subunits, each binding to PDEa or PDEß. Activation of ... More
Structural and Functional Characterization of the Bacterial Type III Secretion Export Apparatus.
AuthorsDietsche T, Tesfazgi Mebrhatu M, Brunner MJ, Abrusci P, Yan J, Franz-Wachtel M, Schärfe C, Zilkenat S, Grin I, Galán JE, Kohlbacher O, Lea S, Macek B, Marlovits TC, Robinson CV, Wagner S
JournalPLoS Pathog
PubMed ID27977800
'Bacterial type III protein secretion systems inject effector proteins into eukaryotic host cells in order to promote survival and colonization of Gram-negative pathogens and symbionts. Secretion across the bacterial cell envelope and injection into host cells is facilitated by a so-called injectisome. Its small hydrophobic export apparatus components SpaP and ... More
New drugs for pharmacological extension of replicative life span in normal and progeroid cells.
AuthorsVatolin S, Radivoyevitch T, Maciejewski JP
JournalNPJ Aging Mech Dis
PubMed ID30675378
A high-throughput anti-aging drug screen was developed that simultaneously measures senescence-associated ß-galactosidase activity and proliferation. Applied to replicatively pre-aged fibroblasts, this screen yielded violuric acid (VA) and 1-naphthoquinone-2-monoxime (N2N1) as its top two hits. These lead compounds extended the replicative life spans of normal and progeroid human cells in a ... More
Structure-based Design of JOC-x, a Conjugatable Tumor Tight Junction Opener to Enhance Cancer Therapy.
AuthorsPitner R, Kim J, Davis-Bergthold J, Turner C, Vassal-Stermann E, Wang H, Adams J, Carter L, Ahlgren JA, Fender P, Lieber A, Carter D, Gray SA
JournalSci Rep
PubMed ID30992466
Disorganized intercellular junctions are critical for maintaining the integrity of solid epithelial tumors and prevent the infiltration of oncological therapies into the bulk of the malignancy. We have developed small, recombinant proteins which bind a critical junction protein, desmoglein 2, triggering the transient and specific opening of tumor tight junctions ... More