Physicochemical studies on the interaction of pancreatic phospholipase A2 with a micellar substrate analog

Jd, Hille; Kelder, Donné-Op den; Sauve, Paul; Gh, de Haas; Mr, Egmond

doi:10.1021/bi00517a019

Cited by 86 publications

(67 citation statements)

References 13 publications

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“…Under these conditions the molecular area of apocytochrome c is 890 + 91 A2/molecule. According to the binding model described by Hille et al [25], assuming independent and identical binding sites, the stoichiometry and K a of apocy- …”

Section: Resultsmentioning

confidence: 99%

Interactions of mitochondrial precursor protein apocytochrome c with phosphatidylserine in model membranes. A monolayer study

Pilon¹,

Jordi²,

Kruijff³

et al. 1987

Biochimica et Biophysica Acta (BBA) - Biomembranes

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(1) The interaction of apocytochrome c with different molecular species of phosphatidyiserine was studied using monolayers at constant surface area or constant surface pressure. The protein inserted readily into dioleoylphosphatidylserine monolayers up to a limiting pressure of 50 raN/m, whereas the interaction decreased with increasing molecular packing of the phosphatidyiserine species, indicating the importance of the hydrophobic core of the lipid layer for the interaction. (2) The high affinity of apocytochrome c for dioleoylphosphatidyiserine is indicated by the low K~ of 0.017/zM. There is little or no interaction with phosphatidylcholines. The importance of charge interactions is underlined by its ionic strength and pH dependency. (3) Experiments using ~4C-labelled apocytochrome c indicate that cholesterol can enhance the protein binding. (4) It was demonstrated that apocytochrome c monomers penetrate the monolayer whereas oligomers can be formed in an adsorbed layer and washed off without changing the surface pressure. Preincubation of apocytochrome c in 3 M guanidine, to obtain the monomeric form, was essential to measure the full effect of interracial interaction. (5) The molecular area of apocytochrome c changed from 1200-1300 .AZ/molecule in the absence of lipid to 700-900 ,A2/molecule after penetration of dioleoyiphosphatidylserine monolayers. (6) Apocytochrome c-dioleoylphosphatidyiserine interactions are only possible when the monolayer is approached from the subphase. It is concluded that the charge interactions are required for binding and penetration of the protein.

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Section: Resultsmentioning

confidence: 99%

Interactions of mitochondrial precursor protein apocytochrome c with phosphatidylserine in model membranes. A monolayer study

Pilon¹,

Jordi²,

Kruijff³

et al. 1987

Biochimica et Biophysica Acta (BBA) - Biomembranes

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“…The binding of pancreatic PLA at the lipid-water interface has been studied extensively using a variety of techniques including gel f'fltration, spectroscopy and hydrodynamic measurements [13][14][15]. When detergent-like n-alkylphosphocholines are used as substrate analogs the stoichiometry of the lipid-protein complex formed between PLA and the zwitterionic detergent micelles varies from 30--40 detergent molecules per protein molecule with 2-3 proteins per particle, and hydrophobic interaction between protein and lipids has been implicated as the main stabilizing factor [14].…”

Section: Discussionmentioning

confidence: 99%

Evidence that the zymogen of phospholipase A2 binds to a negatively charged lipid-water interface

Volwerk¹,

Jost²,

Griffith³

1984

Chemistry and Physics of Lipids

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Evidence is presented that the zymogen of porcine pancreatic phospholipase A~ (prophospholipase A2) interacts with a lipid-water interface provided that the interface has a net negative surface charge. Fluorescence spectroscopy and non-equilibrium gel filtration indicate that binding of prophospholipase A 2 (proPLA) to mixed detergent micelles is dependent on the presence of an anionic detergent. Prophospholipase binding is accompanied by a change in the environment of the single tryptophan residue qualitatively similar to that observed when the active enzyme, phospholipase A s (PLA), binds to micelles. In addition, the rate of tryptic activation of prophospholipase is significantly reduced in the presence of negatively-charged mixed micelles, whereas no change in rate occurs when neutral micelles are present. These observations suggest that the lack of catalytic activity of the zymogen toward organized substrates carrying a negative surface charge cannot be explained by a failure to bind at the lipidwater interface.

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“…The data on the shift of the fluorescence maximum were analysed with a nonlinear regression program using the equation Ko [18].…”

Section: Methodsmentioning

confidence: 99%

Interaction of melittin with negatively charged phospholipids: Consequences for lipid organization

et al. 1987

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A characterization of the structural alterations induced by melittin in model-membranes of dioleoylphosphatidic acid and egg phosphatidylglycerol is presented, based on the use ofaxp-NMR, freeze-fracture electron microscopy and small angle X-ray scattering. In accordance with earlier findings on the cardiolipinmelittin system, melittin is found to have an inverted phase inducing effect on these negatively charged lipids, in contrast to the influence on zwitterionic phospholipids. In phosphatidic acid this is expressed in the formation of an HH phase; in phosphatidylglycerol a less ordered, non-lamellar structure with low water content is adopted.Melittin; Hexagonal HII phase; Protein-lipid interaction; 31p-NMR; Nonbilayer lipid structure; Freeze-fracture electron microscopy

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Physicochemical studies on the interaction of pancreatic phospholipase A2 with a micellar substrate analog

Cited by 86 publications

References 13 publications

Interactions of mitochondrial precursor protein apocytochrome c with phosphatidylserine in model membranes. A monolayer study

Interactions of mitochondrial precursor protein apocytochrome c with phosphatidylserine in model membranes. A monolayer study

Evidence that the zymogen of phospholipase A2 binds to a negatively charged lipid-water interface

Interaction of melittin with negatively charged phospholipids: Consequences for lipid organization

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