scholarly journals Targeted Proteomics for Determining Phosphorylation Site-Specific Associations in Cardiovascular Disease

Circulation ◽  
2012 ◽  
Vol 126 (15) ◽  
pp. 1803-1807 ◽  
Author(s):  
Stuart J. Cordwell ◽  
Melanie Y. White
Keyword(s):  
Cardiovascular Disease ◽  
Phosphorylation Site ◽  
Targeted Proteomics ◽  
Site Specific

An Enzyme-Linked Immunosorbent Assay for Protein Kinase D Activity Using Phosphorylation Site-Specific Antibodies

2007 ◽  
Vol 5 (5) ◽  
pp. 637-644 ◽  
Author(s):  
An Rykx ◽  
Sadia Vancauwenbergh ◽  
Line De Kimpe ◽  
Katrien Janssens ◽  
Sandy Vandoninck ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Immunosorbent Assay ◽  
Phosphorylation Site ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein Kinase D ◽  
Site Specific ◽  
Specific Antibodies

scholarly journals Characterization of Protein Kinase A and Protein Kinase C Phosphorylation of theN-Methyl-D-aspartate Receptor NR1 Subunit Using Phosphorylation Site-specific Antibodies

1997 ◽  
Vol 272 (8) ◽  
pp. 5157-5166 ◽  
Author(s):  
Whittemore G. Tingley ◽  
Michael D. Ehlers ◽  
Kimihiko Kameyama ◽  
Carol Doherty ◽  
Janine B. Ptak ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Protein Kinase A ◽  
Phosphorylation Site ◽  
Site Specific ◽  
Specific Antibodies ◽  
Kinase C ◽  
Aspartate Receptor ◽  
Kinase A

scholarly journals Phosphorylation of the Oncofetal Variant of the Human Bile Salt-dependent Lipase

2001 ◽  
Vol 276 (15) ◽  
pp. 12356-12361 ◽  
Author(s):  
Alain Verine ◽  
Josette Le Petit-Thevenin ◽  
Laurence Panicot-Dubois ◽  
Annick Valette ◽  
Dominique Lombardo
Keyword(s):  
High Pressure ◽  
Bile Salt ◽  
Cho Cells ◽  
Phosphorylation Site ◽  
Chinese Hamster ◽  
Human Bile ◽  
Site Specific ◽  
Phosphorylation Motif ◽  
Extracellular Compartment ◽  
First Time

In this paper, we report, for the first time, the localization of the phosphorylation site of the fetoacinar pancreatic protein (FAPP), which is an oncofetal variant of the pancreatic bile salt-dependent lipase. Using Chinese hamster ovary (CHO) cells transfected with the cDNA encoding FAPP, we radiolabeled the enzyme with32P, and then the protein was purified by affinity chromatography on cholate-immobilized Sepharose column and submitted to a CNBr hydrolysis. Analysis of peptides by high pressure liquid chromatography, associated with the radioactivity profile, revealed that the phosphorylation site is located at threonine 340. Site-specific mutagenesis experiments, in which the threonine was replaced by an alanine residue, were used to invalidate the phosphorylation of FAPP and to study the influence of the modification on the activity and secretion of the enzyme. These studies showed that CHO cells, transfected with the mutated cDNA of FAPP, kept all of their ability to synthesize the protein, but the loss of the phosphorylation motif prevented the release of the protein in the extracellular compartment. However, the mutated enzyme, which was sequestrated in the transfected CHO cells, remains active on bile salt-dependent lipase substrates.

scholarly journals Observation of phosphorylation site-specific dissociation of singly protonated phosphopeptides

2010 ◽  
Vol 21 (1) ◽  
pp. 53-59 ◽  
Author(s):  
Young Sik Shin ◽  
Jeong Hee Moon ◽  
Myung Soo Kim
Keyword(s):  
Phosphorylation Site ◽  
Site Specific

scholarly journals Synthetic-evolution reveals that phosphoregulation of the mitotic kinesin-5 Cin8 is constrained

2018 ◽  
Author(s):  
Alina Goldstein ◽  
Darya Goldman ◽  
Ervin Valk ◽  
Mart Loog ◽  
Liam J. Holt ◽  
...  
Keyword(s):  
Amino Acid ◽  
Phosphorylation Site ◽  
Motor Domain ◽  
Single Amino Acid ◽  
Amino Acid Substitutions ◽  
Phosphorylation Sites ◽  
Wild Type ◽  
Site Specific ◽  
Precise Positioning ◽  
Optimal Regulation

AbstractCdk1 has been found to phosphorylate the majority of its substrates in disordered regions. These phosphorylation sites do not appear to require precise positioning for their function. The mitotic kinesin-5 Cin8 was shown to be phosphoregulated at three Cdk1 sites in disordered loops within its catalytic motor domain. Here, we examined the flexibility of Cin8 phosphoregulation by analyzing the phenotypes of synthetic Cdk1-sites that were systematically generated by single amino-acid substitutions, starting from a phosphodeficient variant of Cin8. Out of 29 synthetic Cdk1 sites that we created, eight were non-functional; 19 were neutral, similar to the phosphodeficient variant; and two gave rise to phosphorylation-dependent spindle phenotypes. Of these two, one site resulted in novel phosphoregulation, and only one site, immediately adjacent to a native Cdk1 site, produced phosphoregulation similar to wild-type. This study shows that, while the gain of a single phosphorylation site can confer regulation and modulate the dynamics of the spindle, to achieve optimal regulation of a mitotic kinesin-5 motor protein, phosphoregulation has to be site-specific and precise.

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