Side chain dynamics of two aromatic amino acids in pancreatic phospholipase A2 as studied by deuterium nuclear magnetic resonance

Biochemistry ◽  
1985 ◽  
Vol 24 (13) ◽  
pp. 3268-3273 ◽  
Author(s):  
P. R. Allegrini ◽  
G. J. M. Van Scharrenburg ◽  
A. J. Slotboom ◽  
G. H. De Haas ◽  
J. Seelig
Keyword(s):  
Amino Acids ◽  
Nuclear Magnetic Resonance ◽  
Magnetic Resonance ◽  
Phospholipase A2 ◽  
Aromatic Amino Acids ◽  
Side Chain ◽  
Chain Dynamics ◽  
Pancreatic Phospholipase ◽  
Pancreatic Phospholipase A2 ◽  
Side Chain Dynamics

The Pyoverdins of Pseudomonas syringae and Pseudomonas cichorii

2004 ◽  
Vol 59 (9-10) ◽  
pp. 613-618 ◽  
Author(s):  
Alain Bultreys ◽  
Isabelle Gheysen ◽  
Mathias Schäfer ◽  
Herbert Budzikiewicz ◽  
Bernard Wathelet
Keyword(s):  
Mass Spectrometry ◽  
Amino Acids ◽  
Nuclear Magnetic Resonance ◽  
Magnetic Resonance ◽  
Pseudomonas Syringae ◽  
Structure Elucidation ◽  
Side Chain ◽  
Closely Related Species ◽  
Pseudomonas Cichorii

Abstract The structure elucidation of the cyclic (lactonic) forms of the pyoverdins with a succinamide side chain originally produced by the closely related species Pseudomonas syringae and P. cichorii is reported. Mass spectrometry and nuclear magnetic resonance analyses as well as the determination of the configuration of the amino acids after degradation indicate that these two pyoverdins differ only by the replacement of the first in-chain serine by glycine. The pyoverdins of P. syringae and P. cichorii and the dihydropyoverdin of P. syringae can be used by both species as siderophores.

19F nuclear magnetic resonance studies of the interaction of inhibitors with chymotrypsin. Ring-fluorinated derivatives of N-trifluoroacetylphenylalanine

1978 ◽  
Vol 31 (10) ◽  
pp. 2179 ◽  
Author(s):  
M Tsavalos ◽  
BC Nicholson ◽  
TM Spotswood
Keyword(s):  
Amino Acids ◽  
Nuclear Magnetic Resonance ◽  
Magnetic Resonance ◽  
Chemical Shift ◽  
Active Site ◽  
Enzyme Inhibitor ◽  
Aromatic Amino Acids ◽  
Binding Parameters ◽  
Magnetic Resonance Studies ◽  
Derivatives Of

The complexes formed between chymotrypsin and the doubly fluorine- labelled inhibitors, o-, m- and p-fluoro-N-trifluoroacetyl-D- phenylalanine and 2,4-difluoro-N-trifluoroacetyl-D-phenylalanine, have been characterized by 19F N.M.R. spectroscopy and binding parameters, ΔB and KI, have been derived. The results confirm the importance of the amido binding site in orienting aromatic amino acids at the active site of chymotrypsin. Changes in ΔB, the change in chemical shift of the enzyme-bound inhibitor, are shown to be a very sensitive probe of enzyme-inhibitor interactions.

Targeting of Porcine Pancreatic Phospholipase A2 to Human Platelets: Introduction of an RGD Sequence by Genetic Engineering

1995 ◽  
Vol 74 (04) ◽  
pp. 1138-1144 ◽  
Author(s):  
A C A P A Bekkers ◽  
H van der Vuurst ◽  
G van Willigen ◽  
J W N Akkerman ◽  
H M Verheij
Keyword(s):  
Amino Acids ◽  
Phospholipase A2 ◽  
Hydrolytic Activity ◽  
Human Platelets ◽  
Rgd Sequence ◽  
Pancreatic Phospholipase ◽  
Pancreatic Phospholipase A2 ◽  
Activated Platelets ◽  
C Terminus ◽  
Platelet Receptor

SummaryThe possibility to induce specific disruption of activated platelets by binding of porcine pancreatic phospholipase A2 (PLA2) was tested by constructing a set of PLA2-mutants containing an Arg-Gly-Asp (RGD) sequence. One mutant was made with RGD as part of a surface-exposed loop (RGDloop). Four mutants were made with RGD as part of a C-terminal extension: one with RGD directly coupled to the C-terminus (RGDc) and three mutants (CRSx) with x = 22,42 and 82 hydrophylic non-charged amino acids between RGD and the enzyme. All mutants retained 20-80% activity of native PLA2 and showed little binding to resting platelets. The binding of the native enzyme and RGDloop was not increased following stimulation. In contrast, the mutants RGDc and CRSx showed stimulation-dependent binding to the platelet receptor GPIIb/IIIa, since GRGDS-peptide and a monoclonal antibody against the complex interfered with binding. In α-thrombin-stimulated platelets, CRS42 and CRS82 induced about 5% hydrolysis of [3H]-arachidonic acid-labeled phospholipids. Stimulation with a combination of a-thrombin and collagen (known to expose phosphatidylserine) increased hydrolysis to 11%. Despite the membrane disruption, the cells did not leak lactate dehydrogenase. We conclude that PLA2 can be targeted to activated platelets by introducing RGD in a C-terminal extension with a minimum distance (42 amino acids) between RGD and the enzyme. However, more hydrolytic activity is required to eliminate activated platelets among a suspension of resting platelets and other blood cells.

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