Determination of Posttranslational Modifications of Photoreceptor Differentiation Factor NRL: Focus on SUMOylation

2012 ◽  
pp. 353-361
Author(s):  
Jerome E. Roger ◽  
Jacob Nellissery ◽  
Anand Swaroop
Keyword(s):  
Posttranslational Modifications ◽  
Differentiation Factor

Determination of Posttranslational Modifications by 2D PAGE: Applications to Polyamines

2017 ◽  
pp. 337-341
Author(s):  
Marta Bitrián ◽  
Antonio F. Tiburcio ◽  
Rubén Alcázar
Keyword(s):  
Posttranslational Modifications ◽  
2D Page

scholarly journals Entianin, a Novel Subtilin-Like Lantibiotic fromBacillus subtilissubsp.spizizeniiDSM 15029Twith High Antimicrobial Activity

2011 ◽  
Vol 77 (5) ◽  
pp. 1698-1707 ◽  
Author(s):  
Sebastian W. Fuchs ◽  
Thorsten W. Jaskolla ◽  
Sophie Bochmann ◽  
Peter Kötter ◽  
Thomas Wichelhaus ◽  
...  
Keyword(s):  
Amino Acid ◽  
Posttranslational Modifications ◽  
Molecular Mechanisms ◽  
Antibiotic Activity ◽  
The Novel ◽  
High Antimicrobial Activity ◽  
Envelope Stress ◽  
Reporter Systems ◽  
First Time

ABSTRACTLantibiotics, such as nisin and subtilin, are lanthionine-containing peptides that exhibit antimicrobial as well as pheromone-like autoinducing activity. Autoinduction is specific for each lantibiotic, and reporter systems for nisin and subtilin autoinduction are available. In this report, we used the previously reported subtilin autoinduction bioassay in combination with mass spectrometric analyses to identify the novel subtilin-like lantibiotic entianin fromBacillus subtilissubsp.spizizeniiDSM 15029T. Linearization of entianin using Raney nickel-catalyzed reductive cleavage enabled, for the first time, the use of tandem mass spectrometry for the fast and efficient determination of an entire lantibiotic primary structure, including posttranslational modifications. The amino acid sequence determined was verified by DNA sequencing of theetnSstructural gene, which confirmed that entianin differs from subtilin at 3 amino acid positions. In contrast toB. subtilisATCC 6633, which produces only small amounts of unsuccinylated subtilin,B. subtilisDSM 15029Tsecretes considerable amounts of unsuccinylated entianin. Entianin was very active against several Gram-positive pathogens, such asStaphylococcus aureusandEnterococcus faecalis.The growth-inhibiting activity of succinylated entianin (S-entianin) was much lower than that of unsuccinylated entianin: a 40-fold higher concentration was required for inhibition. For succinylated subtilin (S-subtilin), a concentration 100-fold higher than that of unsuccinylated entianin was required to inhibit the growth of aB. subtilistest strain. This finding was in accordance with a strongly reduced sensing of cellular envelope stress provided by S-entianin relative to that of entianin. Remarkably, S-entianin and S-subtilin showed considerable autoinduction activity, clearly demonstrating that autoinduction and antibiotic activity underlie different molecular mechanisms.

scholarly journals Position-Specific Analysis and Prediction of Protein Pupylation Sites Based on Multiple Features

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
Xiaowei Zhao ◽  
Jiangyan Dai ◽  
Qiao Ning ◽  
Zhiqiang Ma ◽  
Minghao Yin ◽  
...  
Keyword(s):  
Posttranslational Modifications ◽  
Computational Prediction ◽  
Support Vector ◽  
Good Prediction ◽  
Accurate Identification ◽  
Multiple Features ◽  
Specific Analysis ◽  
Experimental Approaches ◽  
Optimal Feature

Pupylation is one of the most important posttranslational modifications of proteins; accurate identification of pupylation sites will facilitate the understanding of the molecular mechanism of pupylation. Besides the conventional experimental approaches, computational prediction of pupylation sites is much desirable for their convenience and fast speed. In this study, we developed a novel predictor to predict the pupylation sites. First, the maximum relevance minimum redundancy (mRMR) and incremental feature selection methods were made on five kinds of features to select the optimal feature set. Then the prediction model was built based on the optimal feature set with the assistant of the support vector machine algorithm. As a result, the overall jackknife success rate by the new predictor on a newly constructed benchmark dataset was 0.764, and the Mathews correlation coefficient was 0.522, indicating a good prediction. Feature analysis showed that all features types contributed to the prediction of protein pupylation sites. Further site-specific features analysis revealed that the features of sites surrounding the central lysine contributed more to the determination of pupylation sites than the other sites.

scholarly journals Forced Self-Modification Assays as a Strategy to Screen MonoPARP Enzymes

2019 ◽  
Vol 25 (3) ◽  
pp. 241-252
Author(s):  
Tim J. Wigle ◽  
W. David Church ◽  
Christina R. Majer ◽  
Kerren K. Swinger ◽  
Demet Aybar ◽  
...  
Keyword(s):  
Posttranslational Modifications ◽  
Nicotinamide Adenine Dinucleotide ◽  
Chemical Probes ◽  
Fluorescence Immunoassay ◽  
Enzymatic Assays ◽  
Adp Ribosylation ◽  
Biochemical Assays ◽  
Approved Drugs

Mono(ADP-ribosylation) (MARylation) and poly(ADP-ribosylation) (PARylation) are posttranslational modifications found on multiple amino acids. There are 12 enzymatically active mono(ADP-ribose) polymerase (monoPARP) enzymes and 4 enzymatically active poly(ADP-ribose) polymerase (polyPARP) enzymes that use nicotinamide adenine dinucleotide (NAD+) as the ADP-ribose donating substrate to generate these modifications. While there are approved drugs and clinical trials ongoing for the enzymes that perform PARylation, MARylation is gaining recognition for its role in immune function, inflammation, and cancer. However, there is a lack of chemical probes to study the function of monoPARPs in cells and in vivo. An important first step to generating chemical probes for monoPARPs is to develop biochemical assays to enable hit finding, and determination of the potency and selectivity of inhibitors. Complicating the development of enzymatic assays is that it is poorly understood how monoPARPs engage their substrates. To overcome this, we have developed a family-wide approach to developing robust high-throughput monoPARP assays where the enzymes are immobilized and forced to self-modify using biotinylated-NAD+, which is detected using a dissociation-enhanced lanthanide fluorescence immunoassay (DELFIA) readout. Herein we describe the development of assays for 12 monoPARPs and 3 polyPARPs and apply them to understand the potency and selectivity of a focused library of inhibitors across this family.

Determination of posttranslational modifications by mass spectrometry

1992 ◽  
Vol 11 (4) ◽  
pp. 374-375 ◽  
Author(s):  
Toshifumi Takao ◽  
Yasutsugu Shimonishi
Keyword(s):  
Mass Spectrometry ◽  
Posttranslational Modifications

scholarly journals Values of alkaline phosphathase and their isoenzyme profiles in patients with cancer in respect to bone and liver metastasis

2013 ◽  
Vol 21 (1) ◽  
pp. 14-16 ◽  
Author(s):  
Marina Ðokic-Lisanin ◽  
Vesna Pantovic ◽  
Zorica Jovanovic ◽  
Goran Samardzic ◽  
Vladimir Jurisic
Keyword(s):  
Liver Metastasis ◽  
Liver Metastases ◽  
Cancer Patients ◽  
Posttranslational Modifications ◽  
Molecular Forms ◽  
Heat Inactivation ◽  
Healthy Controls ◽  
Alp Activity ◽  
Phosphorus Ester

Background: Alkaline phosphatase is a glycoprotein that catalyzes two kinds of chemical reactions: hydrolysis of phosphorus ester breaking P-O bonds and phospho-transfer reactions in which phosphoric group is transferred to an acceptor molecule. In the human body, ALP exists in multiple molecular forms whose heterogeneity is partly due to genetic factors and partly to posttranslational modifications. The aim was to evaluate a total ALP activity and its isoforms in cancer patients with bone and liver metastasis in comparison to healthy controls. Methods: Human serum was collected from 20 healthy individuals, and 20 cancer patients with bone and liver metastases, with metastases confirmed by ultrasound, computerized tomography and a radiology scan. Determination of ALP was done by the endpoint spectrophotometric method. Isoenzymes were determined by heat inactivation method. Results: In cancer patients, the total ALP activity was significantly higher (p< 0.05) compared to healthy controls. In the sera of cancer patients with liver metastases, the remaining ALP activity was two-fold higher in comparison to bone metastases. Conclusion: Determination of ALP isoenzymes is important but a correct clinical interpretation in the context of other analyses is vital for a proper diagnosis of a disease.

scholarly journals Phosphorylation of Bone Morphogenetic Protein-15 and Growth and Differentiation Factor-9 Plays a Critical Role in Determining Agonistic or Antagonistic Functions

Endocrinology ◽  
2007 ◽  
Vol 149 (2) ◽  
pp. 812-817 ◽  
Author(s):  
Heather E. McMahon ◽  
Shweta Sharma ◽  
Shunichi Shimasaki
Keyword(s):  
Growth Factors ◽  
Bone Morphogenetic Protein ◽  
Posttranslational Modifications ◽  
Critical Role ◽  
Direct Effects ◽  
Phosphorylation State ◽  
Differentiation Factor ◽  
Morphogenetic Protein ◽  
Bone Morphogenetic Protein 15 ◽  
The Impact

Two highly homologous oocyte-secreted growth factors, bone morphogenetic protein (BMP)-15 and growth and differentiation factor (GDF)-9, are known to control folliculogenesis and ovulation through direct effects on granulosa cells in the developing follicles. Although much is known about the expression and biology of these proteins, the impact of posttranslational modifications of BMP-15 and GDF-9 is unknown. Here, we report that: 1) recombinant human (rh) BMP-15 and rhGDF-9 are phosphorylated; 2) the phosphorylation is essential for bioactivity; and 3) the dephosphorylated forms of rhBMP-15 and rhGDF-9 can abolish the bioactivity of rhBMP-15, rhGDF-9, and rhBMP-7, but not rh activin A. These results indicate that the phosphorylation state of rhBMP-15 and rhGDF-9 is a determinant of their agonistic and antagonistic activities.

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