Surface topology of Minibody by selective chemical modifications and mass spectrometry

The surface topology of the Minibody, a small de novo‐designed β‐protein, has been probed by a strategy that combines selective chemical modification with a variety of reagents and mass spectrometric analysis of the modified fragments. Under appropriate conditions, the susceptibility of individual r...

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Bibliographic Details
Published in:Protein science 1997-09, Vol.6 (9), p.1901-1909
Main Authors: Zappacosta, Francesca, Ingallinella, Paolo, Scaloni, Andrea, Pessi, Antonello, Bianchi, Elisabetta, Sollazzo, Maurizio, Tramontano, Anna, Marino, Gennaro, Pucci, Piero
Format: Article
Language:English
Subjects:
Acetylation
Amino Acid Sequence
Arginine - chemistry
Carrier Proteins - chemistry
Chromatography, High Pressure Liquid
Hydrolysis
Immunoglobulins - chemistry
Lysine - chemistry
Mass Spectrometry
Minibody
Molecular Sequence Data
Molecular Weight
Peptide Fragments - chemistry
Protein Structure, Secondary
protein surface topology
Recombinant Proteins - chemistry
selective chemical modifications
Tyrosine - chemistry
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Summary:The surface topology of the Minibody, a small de novo‐designed β‐protein, has been probed by a strategy that combines selective chemical modification with a variety of reagents and mass spectrometric analysis of the modified fragments. Under appropriate conditions, the susceptibility of individual residues primarily depends on their surface accessibility so that their relative reactivities can be correlated with their position in the tertiary structure of the protein. Moreover, this approach provides information on interacting residues, since intramolecular interactions might greatly affect the reactivity of individual side chains by altering their pKa values. The results of this study indicate that, while overall the Minibody model is correct, the β‐sheet formed by the N‐ and C‐terminal segments is most likely distorted. This is also in agreement with previous results that were obtained using a similar approach where mass spectrometry was used to identify Minibody fragments from limited proteolysis (Zappacosta F, Pessi A, Bianchi E, Venturini s, Sollazzo M, Tramontano A, Marino G, Pucci P. 1996. Probing the tertiary structure of proteins by limited proteolysis and mass spectrometry: The case of Minibody. Protein Sci 5902‐813). The chemical modification approach, in combination with limited proteolysis procedures, can provide useful, albeit partial, structural information to complement simulation techniques. This is especially valuable when, as in the Minibody case, an NMR and/or X‐ray structure cannot be obtained due to insufficient solubility of the molecule.
ISSN:0961-8368
1469-896X
DOI:10.1002/pro.5560060911