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Title of Journal: Protein J

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Abbravation: The Protein Journal

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Kluwer Academic Publishers-Plenum Publishers

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DOI

10.1016/0148-9062(85)90201-3

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1573-4943

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Biological and Structural Characterization of Crot

Authors: Luis Alberto PonceSoto Bruno Lomonte Lea RodriguesSimioni José Camillo Novello Sergio Marangoni
Publish Date: 2007/01/03
Volume: 26, Issue: 4, Pages: 221-230
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Abstract

A new crotoxin B isoform PLA2 F6a from Crotalus durissus collilineatus was purified from by one step reverse phase HPLC chromatography using μBondapack C18 column analytic The new crotoxin B isoform PLA2 F6a complex crotoxin the catalytic subunit crotoxin B isoform PLA2 F6a and two crotapotin isoforms F3 and F4 were isolated from the venom of Crotalus durissus collilineatus The crotapotins isoforms F3 and F4 had similar chemical properties the two proteins different in their ability to inhibit of isoforms of PLA2 F6 and F6a The molecular masses estimated by MALDITOF mass spectrometry were crotoxin B 1494314 Da crotapotin F3 869324 Da and crotapotin F4 9 31456 Da The new crotoxin B isoform PLA2 F6a contained 122 amino acid residues and a pI of 858 Its amino acid sequence presents high identity with those of other PLA2s particularly in the calcium binding loop and active site helix 3 It also presents similarities in the Cterminal region with other myotoxic PLA2s The new crotoxin B isoform PLA2 F6a contained 122 amino acid residues with a primary structure of HLLQFNKMIK FETRRNAIPP YAFYGCYCGW GGRGRPKDAT DRCCFVHDCC YGKLAKCNTK WDFYRYSLKS GYITCGKGTW CEEQICECDR VAAECLRRSL STYRYGYMIY PDSRCRGPSE TC A neuromuscular blocking activity was induced by crotoxin and new crotoxin B isoform PLA2 F6a in the isolated mouse phrenic nerve diaphragm and the biventer cervicis chick nervemuscle preparation Whole crotoxin was devoid of cytolytic activity upon myoblasts and myotubes in vitro whereas new crotoxin B isoform PLA2 F6a was clearly cytotoxic to these cells

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  2. The Use of the Free Metal – Temperature ‘Phase Diagrams’ for Studies of Single Site Metal Binding Proteins
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  4. Effects of Pulsed Electric Field (PEF) Treatment on Enhancing Activity and Conformation of α-Amylase
  5. Isolation, Purification and Characterization of Two Laccases from Carrot ( Daucus carota L.) and Their Response to Abiotic and Metal Ions Stresses
  6. Conformational Change in the Active Site of Streptococcal Unsaturated Glucuronyl Hydrolase Through Site-Directed Mutagenesis at Asp-115
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  8. Biophysical Analysis of the N-Terminal Domain from the Human Protein Phosphatase 1 Nuclear Targeting Subunit PNUTS Suggests an Extended Transcription Factor TFIIS-Like Fold
  9. Improved Protein Overexpression and Purification Strategies for Structural Studies of Cyanobacterial Metal-Responsive Transcription Factor, SmtB from Marine Synechococcus sp. PCC 7002
  10. Influence of Polyols on the Stability and Kinetic Parameters of Invertase from Candida utilis : Correlation with the Conformational Stability and Activity
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  12. The Role of Strong Electrostatic Interactions at the Dimer Interface of Human Glutathione Synthetase
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