1qm6: Difference between revisions

Revision as of 15:41, 21 February 2008

R32 FORM OF CLOSTRIDIUM PERFRINGENS ALPHA-TOXIN STRAIN

OverviewOverview

Alpha-toxin is the key determinant in gas-gangrene. The toxin, a phospholipase C, cleaves phosphatidylcholine in eukaryotic cell membranes. Calcium ions have been shown to be required for the specific binding of toxin to membranes prior to phospholipid cleavage. Reported X-ray crystallographic structures of the toxin show that the C-terminal domain has a fold that is analogous to the eukaryotic calcium and membrane-binding C2 domains. We report the binding sites for three calcium ions that have been identified, by crystallographic methods, in the C-terminal domain of the protein close to the postulated membrane-binding surface. The position of these ions at the tip of the domain, and their function (to facilitate membrane binding) is similar to that of calcium ions observed bound to C2 domains. Using the optical spectroscopic techniques of circular dichroism (CD) and fluorescence spectroscopy, pronounced changes to both near and far-UV CD and tryptophan emission fluorescence upon addition of calcium to the C-terminal domain of alpha-toxin have been observed. The changes in near-UV CD, fluorescence enhancement and a 2 nm blue-shift in the fluorescence emission spectrum are consistent with tryptophan residue(s) becoming more immobilised in a hydrophobic environment. Calcium binding appears to be low-affinity: Kd approximately 175-250 microM at pH 8 assuming a 1:1 stoichiometry. as measured by spectroscopic methods.

About this StructureAbout this Structure

1QM6 is a Single protein structure of sequence from Clostridium perfringens with as ligand. Active as Phospholipase C, with EC number 3.1.4.3 Known structural/functional Sites: , , and . Full crystallographic information is available from OCA.

ReferenceReference

Characterisation of the calcium-binding C-terminal domain of Clostridium perfringens alpha-toxin., Naylor CE, Jepson M, Crane DT, Titball RW, Miller J, Basak AK, Bolgiano B, J Mol Biol. 1999 Dec 3;294(3):757-70. PMID:10610794

Page seeded by OCA on Thu Feb 21 14:41:16 2008

Proteopedia Page Contributors and Editors (what is this?)Proteopedia Page Contributors and Editors (what is this?)

OCA

@@ Line 4: / Line 4: @@
 ==Overview==
-Alpha-toxin is the key determinant in gas-gangrene. The toxin, a, phospholipase C, cleaves phosphatidylcholine in eukaryotic cell membranes., Calcium ions have been shown to be required for the specific binding of, toxin to membranes prior to phospholipid cleavage. Reported X-ray, crystallographic structures of the toxin show that the C-terminal domain, has a fold that is analogous to the eukaryotic calcium and, membrane-binding C2 domains. We report the binding sites for three calcium, ions that have been identified, by crystallographic methods, in the, C-terminal domain of the protein close to the postulated membrane-binding, surface. The position of these ions at the tip of the domain, and their, function (to facilitate membrane binding) is similar to that of calcium, ions observed bound to C2 domains. Using the optical spectroscopic, techniques of circular dichroism (CD) and fluorescence spectroscopy, pronounced changes to both near and far-UV CD and tryptophan emission, fluorescence upon addition of calcium to the C-terminal domain of, alpha-toxin have been observed. The changes in near-UV CD, fluorescence, enhancement and a 2 nm blue-shift in the fluorescence emission spectrum, are consistent with tryptophan residue(s) becoming more immobilised in a, hydrophobic environment. Calcium binding appears to be low-affinity: Kd, approximately 175-250 microM at pH 8 assuming a 1:1 stoichiometry. as, measured by spectroscopic methods.
+Alpha-toxin is the key determinant in gas-gangrene. The toxin, a phospholipase C, cleaves phosphatidylcholine in eukaryotic cell membranes. Calcium ions have been shown to be required for the specific binding of toxin to membranes prior to phospholipid cleavage. Reported X-ray crystallographic structures of the toxin show that the C-terminal domain has a fold that is analogous to the eukaryotic calcium and membrane-binding C2 domains. We report the binding sites for three calcium ions that have been identified, by crystallographic methods, in the C-terminal domain of the protein close to the postulated membrane-binding surface. The position of these ions at the tip of the domain, and their function (to facilitate membrane binding) is similar to that of calcium ions observed bound to C2 domains. Using the optical spectroscopic techniques of circular dichroism (CD) and fluorescence spectroscopy, pronounced changes to both near and far-UV CD and tryptophan emission fluorescence upon addition of calcium to the C-terminal domain of alpha-toxin have been observed. The changes in near-UV CD, fluorescence enhancement and a 2 nm blue-shift in the fluorescence emission spectrum are consistent with tryptophan residue(s) becoming more immobilised in a hydrophobic environment. Calcium binding appears to be low-affinity: Kd approximately 175-250 microM at pH 8 assuming a 1:1 stoichiometry. as measured by spectroscopic methods.
 ==About this Structure==
@@ Line 14: / Line 14: @@
 [[Category: Phospholipase C]]
 [[Category: Single protein]]
-[[Category: Basak, A.K.]]
+[[Category: Basak, A K.]]
 [[Category: Miller, J.]]
-[[Category: Naylor, C.E.]]
+[[Category: Naylor, C E.]]
-[[Category: Titball, R.W.]]
+[[Category: Titball, R W.]]
 [[Category: ZN]]
 [[Category: c2 domain]]
@@ Line 24: / Line 24: @@
 [[Category: zinc phospholipase c]]
-''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Feb  3 10:01:05 2008''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 14:41:16 2008''