scholarly journals Analysis of protein composition and protein expression in the tear fluid of patients with congenital aniridia

2013 ◽  
Vol 94 ◽  
pp. 78-88 ◽  
Author(s):  
Robert Ihnatko ◽  
Ulla Edén ◽  
Neil Lagali ◽  
Anette Dellby ◽  
Per Fagerholm
Author(s):  
Paul Musille ◽  
Eric Ortlund

The 1.90 Å resolution X-ray crystal structure of glycerol dehydrogenase derived from contaminating bacteria present during routineEscherichia coliprotein expression is presented. This off-target enzyme showed intrinsic affinity for Ni2+-Sepharose, migrated at the expected molecular mass for the target protein during gel filtration and was crystallized before it was realised that contamination had occurred. In this study, it is shown that liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) can efficiently identify the protein composition of crystals in a crystallization experiment as part of a structure-determination pipeline for an unknown protein. The high-resolution X-ray data enabled sequencing directly from the electron-density maps, allowing the source of contamination to be placed within theSerratiagenus. Incorporating additional protein-identity checks, such as tandem LC-MS/MS, earlier in the protein expression, purification and crystallization workflow may have prevented the unintentional structure determination of this metabolic enzyme, which represents the first enterobacterial glycerol dehydrogenase reported to date.


2019 ◽  
Vol 20 (17) ◽  
pp. 4162 ◽  
Author(s):  
Fabia Fricke ◽  
Malwina Michalak ◽  
Uwe Warnken ◽  
Ingrid Hausser ◽  
Martina Schnölzer ◽  
...  

Microsatellite unstable (MSI) colorectal cancers (CRCs) are characterized by mutational inactivation of Transforming Growth Factor Beta Receptor Type 2 (TGFBR2). TGFBR2-deficient CRCs present altered target gene and protein expression. Such cellular alterations modulate the content of CRC-derived extracellular vesicles (EVs). EVs function as couriers of proteins, nucleic acids, and lipids in intercellular communication. At a qualitative level, we have previously shown that TGFBR2 deficiency causes overall alterations in the EV protein content. To deepen the basic understanding of altered protein dynamics, this work aimed to determine TGFBR2-dependent EV protein signatures in a quantitative manner. Using a stable isotope labeling with amino acids in cell culture (SILAC) approach for mass spectrometry-based quantification, 48 TGFBR2-regulated proteins were identified in MSI CRC-derived EVs. Overall, TGFBR2 deficiency caused upregulation of several EV proteins related to the extracellular matrix and nucleosome as well as downregulation of proteasome-associated proteins. The present study emphasizes the general overlap of proteins between EVs and their parental CRC cells but also highlights the impact of TGFBR2 deficiency on EV protein composition. From a clinical perspective, TGFBR2-regulated quantitative differences of protein expression in EVs might nominate novel biomarkers for liquid biopsy-based MSI typing in the future.


2013 ◽  
Vol 91 ◽  
pp. 0-0
Author(s):  
E IOMDINA ◽  
E TARUTTA ◽  
I KURYLEVA ◽  
Y AKSENOVA ◽  
E SURINA ◽  
...  

2010 ◽  
Vol 34 (8) ◽  
pp. S12-S12
Author(s):  
Hong‑Ge Li ◽  
Chen Min Xu ◽  
Kun Li ◽  
Ya Ni ◽  
Wen‑Ying Chen ◽  
...  

2000 ◽  
Vol 111 (4) ◽  
pp. 1118-1121 ◽  
Author(s):  
A. Bellahcene ◽  
I. Van Riet ◽  
C. de Greef ◽  
N. Antoine ◽  
M. F. Young ◽  
...  

2007 ◽  
Vol 177 (4S) ◽  
pp. 222-222
Author(s):  
Mireia Musquera ◽  
Maria J. Ribal ◽  
Yolanda Arce ◽  
Humberto Villavicencio ◽  
Fernando Algaba ◽  
...  

Sign in / Sign up

Export Citation Format

Share Document