Amyloid recognition by serum amyloid P component
Amyloid recognition by serum amyloid P component
The X-ray crystal structures of serum amyloid P component (SAP) with bound (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic acid (Ro 63-8695) and related components have been solved to elucidate the molecular basis of the action of Ro 63-8695, a potential amyloid mobilizing drug for treatment of human amyloid disease. The structure of SAP in the presence of N-acetyl-D-proline has been determined to a resolution of 2.4A using a previous solved SAP structure as the phasing model (unit cell dimensions a=96.1A, b=70.8A, c=103.6A, and b=96.8). The carboxyl group of N-acetyl-D-proline is bound in the double calcium-binding site of each subunit, orientating the pyrrolidine ring into the adjacent hydrophobic pocket formed between Leu6, Tyr64, and Tyr74. The structure of SAP co-crystallised with Ro 63-8695 has been determined to a resolution of 3.2A by molecular replacement (unit cell dimensions a=b=230.9A and c=281.4A). This shows the formation of a ligand-induced decamer, where two SAP pentamers are reversibly cross-linked by five Ro 63-8695 molecules. Binding of the Ro 63-86945 molecule head group shows close supervision with the higher resolution N-acetyl-D-proline structure. The alkyl linker adopts a kinked rotamer about carbons 2 and 3 to facilitate binding of the head groups to the two-fold axis related subunits. The best fit of the electron density is found when both peptide bonds preceding the pyrrolidine ring adopt a cis conformation. Nuclear magnetic resonance spectroscopy has estimated this cis-cis isomer to contribute only ~6% of the Ro 63-8695 population in free solution. SAP has also been found to enhance the refolding yield of denatured lactate dehydrogenase and protects against enzyme inactivation during agitation through a calcium independent site.
University of Southampton
Purvis, Alan
2534bb2c-4b8a-496e-a645-ef4b620551cf
2002
Purvis, Alan
2534bb2c-4b8a-496e-a645-ef4b620551cf
Purvis, Alan
(2002)
Amyloid recognition by serum amyloid P component.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
The X-ray crystal structures of serum amyloid P component (SAP) with bound (R)-1-[6-[(R)-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]-pyrrolidine-2-carboxylic acid (Ro 63-8695) and related components have been solved to elucidate the molecular basis of the action of Ro 63-8695, a potential amyloid mobilizing drug for treatment of human amyloid disease. The structure of SAP in the presence of N-acetyl-D-proline has been determined to a resolution of 2.4A using a previous solved SAP structure as the phasing model (unit cell dimensions a=96.1A, b=70.8A, c=103.6A, and b=96.8). The carboxyl group of N-acetyl-D-proline is bound in the double calcium-binding site of each subunit, orientating the pyrrolidine ring into the adjacent hydrophobic pocket formed between Leu6, Tyr64, and Tyr74. The structure of SAP co-crystallised with Ro 63-8695 has been determined to a resolution of 3.2A by molecular replacement (unit cell dimensions a=b=230.9A and c=281.4A). This shows the formation of a ligand-induced decamer, where two SAP pentamers are reversibly cross-linked by five Ro 63-8695 molecules. Binding of the Ro 63-86945 molecule head group shows close supervision with the higher resolution N-acetyl-D-proline structure. The alkyl linker adopts a kinked rotamer about carbons 2 and 3 to facilitate binding of the head groups to the two-fold axis related subunits. The best fit of the electron density is found when both peptide bonds preceding the pyrrolidine ring adopt a cis conformation. Nuclear magnetic resonance spectroscopy has estimated this cis-cis isomer to contribute only ~6% of the Ro 63-8695 population in free solution. SAP has also been found to enhance the refolding yield of denatured lactate dehydrogenase and protects against enzyme inactivation during agitation through a calcium independent site.
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Published date: 2002
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Local EPrints ID: 464712
URI: http://eprints.soton.ac.uk/id/eprint/464712
PURE UUID: 5d1030e3-0fd3-47c7-8e2c-efce333ebad0
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Date deposited: 04 Jul 2022 23:58
Last modified: 16 Mar 2024 19:43
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Author:
Alan Purvis
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