The University of Southampton
University of Southampton Institutional Repository

Amyloid recognition by serum amyloid P component

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
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.

Text
847352.pdf - Version of Record
Available under License University of Southampton Thesis Licence.
Download (40MB)

More information

Published date: 2002

Identifiers

Local EPrints ID: 464712
URI: http://eprints.soton.ac.uk/id/eprint/464712
PURE UUID: 5d1030e3-0fd3-47c7-8e2c-efce333ebad0

Catalogue record

Date deposited: 04 Jul 2022 23:58
Last modified: 23 Jul 2022 02:14

Export record

Contributors

Author: Alan Purvis

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of http://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×