Assignment of the Complete Disulphide Bridge Pattern in the Human Recombinant Follitropin β -Chain
-
Angela Amoresano
Abstract
The chemical assessment of the complete disulphide bridge pattern in the βchain of human recombinant follicotropin (βFSH) was accomplished by integrating classical biochemical methodologies with mass spectrometric procedures. A proteolytic strategy consisting of a double digestion of native βFSH using the broadspecificity protease subtilisin first, followed by trypsin, was employed. The resulting peptide mixture was directly analysed by FABMS, leading to the assignment of the first three disulphide bridges. The remaining SS bridges were determined by HPLC fractionation of the proteolytic digest followed by ESMS analysis of the individual fractions. The pattern of cysteine couplings in βFSH was determined as: Cys3-Cys51, Cys17-Cys66, Cys20-Cys104, Cys28-Cys82, Cys32-Cys84 and Cys87-Cys94, confirming the arrangement inferred from the crystal structure of the homologous βCG. A subset of the SS bridge pattern comprising Cys3-Cys51, Cys28- Cys82 and Cys32-Cys84 constitutes a cysteine knot motif similar to that found in the growth factor superfamily.
Copyright © 2001 by Walter de Gruyter GmbH & Co. KG
Artikel in diesem Heft
- Biological Activity of Mammalian Transcriptional Repressors
- Cloning and Characterisation of Chlorophyll Synthase from Avena sativa
- DNA Damage by 3,6-Dihydropyrazine-2,5-Dipropanoic Acid, the Cyclic Dimerization Product of 5-Aminolevulinic Acid
- Chicken Erythrocyte Pyrimidine 5-Nucleotidase: Purification and Characterization of the Subclass I Enzyme
- Coenzymes Q9 and Q10, Vitamin E and Peroxidation in Rat Synaptic and Non-Synaptic Occipital Cerebral Cortex Mitochondria during Ageing
- Membrane Activity of (Cys48Ser) Lung Surfactant Protein B Increases with Dimerisation
- Participation of Residue F552 in Domain III of the Protective Antigen in the Biological Activity of Anthrax Lethal Toxin
- Purification of Chrysancorin, a Novel Antifungal Protein with Mitogenic Activity from Garland Chrysanthemum Seeds
- Legumain Forms from Plants and Animals Differ in Their Specificity
- Assignment of the Complete Disulphide Bridge Pattern in the Human Recombinant Follitropin β -Chain
- The Small GTPase Ras Is Involved in Growth Factor-Regulated Expression of the α1 Integrin Subunit in PC12 Cells
- The Mouse Gap Junction Gene Connexin29 Is Highly Expressed in Sciatic Nerve and Regulated during Brain Development
- A Selenocysteine-Containing Peroxiredoxin from the Strictly Anaerobic Organism Eubacterium acidaminophilum
- Characterization of C-Terminally Truncated Human Tissue Inhibitor of Metalloproteinases-4 Expressed in Pichia pastoris
Artikel in diesem Heft
- Biological Activity of Mammalian Transcriptional Repressors
- Cloning and Characterisation of Chlorophyll Synthase from Avena sativa
- DNA Damage by 3,6-Dihydropyrazine-2,5-Dipropanoic Acid, the Cyclic Dimerization Product of 5-Aminolevulinic Acid
- Chicken Erythrocyte Pyrimidine 5-Nucleotidase: Purification and Characterization of the Subclass I Enzyme
- Coenzymes Q9 and Q10, Vitamin E and Peroxidation in Rat Synaptic and Non-Synaptic Occipital Cerebral Cortex Mitochondria during Ageing
- Membrane Activity of (Cys48Ser) Lung Surfactant Protein B Increases with Dimerisation
- Participation of Residue F552 in Domain III of the Protective Antigen in the Biological Activity of Anthrax Lethal Toxin
- Purification of Chrysancorin, a Novel Antifungal Protein with Mitogenic Activity from Garland Chrysanthemum Seeds
- Legumain Forms from Plants and Animals Differ in Their Specificity
- Assignment of the Complete Disulphide Bridge Pattern in the Human Recombinant Follitropin β -Chain
- The Small GTPase Ras Is Involved in Growth Factor-Regulated Expression of the α1 Integrin Subunit in PC12 Cells
- The Mouse Gap Junction Gene Connexin29 Is Highly Expressed in Sciatic Nerve and Regulated during Brain Development
- A Selenocysteine-Containing Peroxiredoxin from the Strictly Anaerobic Organism Eubacterium acidaminophilum
- Characterization of C-Terminally Truncated Human Tissue Inhibitor of Metalloproteinases-4 Expressed in Pichia pastoris
