scholarly journals Insight into the conformational dynamics of specific regions of porcine pancreatic phospholipase A2 from a time-resolved fluorescence study of a genetically inserted single tryptophan residue

Biochemistry ◽  
1991 ◽  
Vol 30 (36) ◽  
pp. 8771-8785 ◽  
Author(s):  
Oscar P. Kuipers ◽  
Michel Vincent ◽  
Jean Claude Brochon ◽  
Hubertus M. Verheij ◽  
Gerard H. De Haas ◽  
...  
Keyword(s):  
Phospholipase A2 ◽  
Conformational Dynamics ◽  
Tryptophan Residue ◽  
Time Resolved Fluorescence ◽  
Fluorescence Study ◽  
Time Resolved ◽  
Pancreatic Phospholipase ◽  
Pancreatic Phospholipase A2 ◽  
Insight Into

Time-resolved fluoroimmunoassay of human pancreatic phospholipase A2.

1983 ◽  
Vol 29 (10) ◽  
pp. 1777-1780 ◽  
Author(s):  
J U Eskola ◽  
T J Nevalainen ◽  
T N Lövgren
Keyword(s):  
Phospholipase A2 ◽  
Solid Phase ◽  
High Sensitivity ◽  
Fluorometric Detection ◽  
Time Resolved ◽  
Antibody Technique ◽  
Pancreatic Phospholipase ◽  
Pancreatic Phospholipase A2 ◽  
Immunoreactive Phospholipase A2 ◽  
Solid Phase Assay

Abstract We describe an immunofluorometric assay for human pancreatic phospholipase A2 based on time-resolved fluorescence. The labeled antibody technique in combination with the time-resolved 1-s fluorometric detection of the europium label, which essentially eliminates all background fluorescence, resulted in a high sensitivity (20 ng/L) and a wide (5000-fold) linear range. Nonspecific binding was minimized by treating the solid-phase antibody with NaSCN before coating, to remove endogenous antigen, and by immunosorbent purification of the antibody before labeling with europium. This is a one-incubation, multi-site, solid-phase assay on polystyrene microtiter strips, even though a polyclonal antibody was used. As measured by this assay, activity of immunoreactive phospholipase A2 was found to be above normal in sera of patients suffering from acute pancreatitis.

scholarly journals Structural Insight into the Activation Mechanism of Human Pancreatic Prophospholipase A2

2009 ◽  
Vol 284 (24) ◽  
pp. 16659-16666 ◽  
Author(s):  
Wei Xu ◽  
Lina Yi ◽  
Yumei Feng ◽  
Ling Chen ◽  
Jinsong Liu
Keyword(s):  
Phospholipase A2 ◽  
Catalytic Triad ◽  
Activation Mechanism ◽  
X Ray ◽  
Pancreatic Phospholipase ◽  
Pancreatic Phospholipase A2 ◽  
Obesity And Diabetes ◽  
Membrane Bound ◽  
Structural Insight ◽  
Insight Into

Pancreatic phospholipase A2 (phospholipase A2 group 1B, G1B) belongs to the superfamily of secreted phospholipase A2 (PLA2) enzymes. G1B has been proposed to be a potential target for diseases such as hypertension, obesity, and diabetes. Human pancreatic prophospholipase A2 (pro-hG1B) is activated by cleavage of the first seven-residue propeptide (phospholipase A2 propeptide, PROP). However, questions still remain on the mode of action for pro-hG1B. In this work, we expressed pro-hG1B in Pichia pastoris and determined the crystal structure at 1.55-Å resolution. The x-ray structure demonstrates that pro-hG1B forms a trimer. In addition, PROP occupies the catalytic cavity and can be self-cleaved at 37 °C. A new membrane-bound surface and activation mechanism are proposed based on the trimeric model of pro-hG1B. We also propose a new autoproteolytic mechanism for pro-hG1B by the reaction triad Asp49-Arg0-Ser(-2) that is similar to the serine protease catalytic triad.

Time-resolved fluorescence study of VU-9 calmodulin, an engineered calmodulin possessing a single tryptophan residue

Biochemistry ◽  
1989 ◽  
Vol 28 (14) ◽  
pp. 6093-6098 ◽  
Author(s):  
M. Chabbert ◽  
Marie Claude Kilhoffer ◽  
D. Martin Watterson ◽  
Jacques Haiech ◽  
H. Lami
Keyword(s):  
Tryptophan Residue ◽  
Time Resolved Fluorescence ◽  
Fluorescence Study ◽  
Time Resolved

Rotational dynamics of the single tryptophan of porcine pancreatic phospholipase A2, its zymogen, and an enzyme/micelle complex. A steady-state and time-resolved anisotropy study

Biochemistry ◽  
1988 ◽  
Vol 27 (17) ◽  
pp. 6618-6628 ◽  
Author(s):  
Richard D. Ludescher ◽  
Iain D. Johnson ◽  
Johannes J. Volwerk ◽  
Gerard H. De Haas ◽  
Patricia C. Jost ◽  
...  
Keyword(s):  
Steady State ◽  
Phospholipase A2 ◽  
Rotational Dynamics ◽  
Time Resolved ◽  
Pancreatic Phospholipase ◽  
Pancreatic Phospholipase A2
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