Preliminary X-ray analysis of cellobiohydrolase Cel7B from Melanocarpus albomyces

Tarja Parkkinen (Corresponding Author), Anu Koivula, Jari Vehmaanperä, Juha Rouvinen

Research output: Contribution to journalArticleScientificpeer-review

4 Citations (Scopus)

Abstract

Enantioselective antibodies can separate the enantiomers of a chiral compound in a highly specific manner. We have recently reported the cloning and applications of a recombinant Fab-fragment, ENA11His, in the enantioseparation of a drug candidate, finrozole, which contains two chiral centers. Here, the crystal structures of this enantioselective antibody Fab-fragment are determined in the absence of the hapten at a resolution of 2.75 Å, and in the presence of the hapten at 2.05 Å resolution. The conformation of the protein was found to be similar in both free and complex forms. The hapten molecule was tightly bound in a deep cleft between the light and heavy chains of the Fab-fragment. The complex structure also allowed us to describe the molecular basis for enantioselectivity and to deduce the absolute configurations of all the four different stereoisomers (a–d) of finrozole. The ENA11His antibody fragment selectively binds the SR (a) enantiomer from the racemic mixture of a and d-enantiomers, thus allowing separation from the pharmacologically most active RS enantiomer (d). In particular, Asp95 and Asn35 of the H-chain in the ENA11 His antibody seem to provide this specificity through hydrogen bonding.
Original languageEnglish
Pages (from-to)754-757
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
VolumeF63
Issue number9
DOIs
Publication statusPublished - 2007
MoE publication typeA1 Journal article-refereed

Fingerprint

Cellulose 1,4-beta-Cellobiosidase
Immunoglobulin Fab Fragments
Enantiomers
Haptens
enantiomers
X ray analysis
antibodies
Immunoglobulin Fragments
fragments
X-Rays
Antibodies
Protein Conformation
Stereoisomerism
x rays
Hydrogen Bonding
Organism Cloning
Enantioselectivity
Cloning
Conformations
Light

Keywords

  • enantioselective antibody
  • Fab-fragment
  • complex
  • crystal structure
  • finrozole

Cite this

@article{f436a0b24cb048d2b6145976ff3da5fc,
title = "Preliminary X-ray analysis of cellobiohydrolase Cel7B from Melanocarpus albomyces",
abstract = "Enantioselective antibodies can separate the enantiomers of a chiral compound in a highly specific manner. We have recently reported the cloning and applications of a recombinant Fab-fragment, ENA11His, in the enantioseparation of a drug candidate, finrozole, which contains two chiral centers. Here, the crystal structures of this enantioselective antibody Fab-fragment are determined in the absence of the hapten at a resolution of 2.75 {\AA}, and in the presence of the hapten at 2.05 {\AA} resolution. The conformation of the protein was found to be similar in both free and complex forms. The hapten molecule was tightly bound in a deep cleft between the light and heavy chains of the Fab-fragment. The complex structure also allowed us to describe the molecular basis for enantioselectivity and to deduce the absolute configurations of all the four different stereoisomers (a–d) of finrozole. The ENA11His antibody fragment selectively binds the SR (a) enantiomer from the racemic mixture of a and d-enantiomers, thus allowing separation from the pharmacologically most active RS enantiomer (d). In particular, Asp95 and Asn35 of the H-chain in the ENA11 His antibody seem to provide this specificity through hydrogen bonding.",
keywords = "enantioselective antibody, Fab-fragment, complex, crystal structure, finrozole",
author = "Tarja Parkkinen and Anu Koivula and Jari Vehmaanper{\"a} and Juha Rouvinen",
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language = "English",
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journal = "Acta Crystallographica Section F: Structural Biology Communications",
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}

Preliminary X-ray analysis of cellobiohydrolase Cel7B from Melanocarpus albomyces. / Parkkinen, Tarja (Corresponding Author); Koivula, Anu; Vehmaanperä, Jari; Rouvinen, Juha.

In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications, Vol. F63, No. 9, 2007, p. 754-757.

Research output: Contribution to journalArticleScientificpeer-review

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T1 - Preliminary X-ray analysis of cellobiohydrolase Cel7B from Melanocarpus albomyces

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AU - Koivula, Anu

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AU - Rouvinen, Juha

PY - 2007

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N2 - Enantioselective antibodies can separate the enantiomers of a chiral compound in a highly specific manner. We have recently reported the cloning and applications of a recombinant Fab-fragment, ENA11His, in the enantioseparation of a drug candidate, finrozole, which contains two chiral centers. Here, the crystal structures of this enantioselective antibody Fab-fragment are determined in the absence of the hapten at a resolution of 2.75 Å, and in the presence of the hapten at 2.05 Å resolution. The conformation of the protein was found to be similar in both free and complex forms. The hapten molecule was tightly bound in a deep cleft between the light and heavy chains of the Fab-fragment. The complex structure also allowed us to describe the molecular basis for enantioselectivity and to deduce the absolute configurations of all the four different stereoisomers (a–d) of finrozole. The ENA11His antibody fragment selectively binds the SR (a) enantiomer from the racemic mixture of a and d-enantiomers, thus allowing separation from the pharmacologically most active RS enantiomer (d). In particular, Asp95 and Asn35 of the H-chain in the ENA11 His antibody seem to provide this specificity through hydrogen bonding.

AB - Enantioselective antibodies can separate the enantiomers of a chiral compound in a highly specific manner. We have recently reported the cloning and applications of a recombinant Fab-fragment, ENA11His, in the enantioseparation of a drug candidate, finrozole, which contains two chiral centers. Here, the crystal structures of this enantioselective antibody Fab-fragment are determined in the absence of the hapten at a resolution of 2.75 Å, and in the presence of the hapten at 2.05 Å resolution. The conformation of the protein was found to be similar in both free and complex forms. The hapten molecule was tightly bound in a deep cleft between the light and heavy chains of the Fab-fragment. The complex structure also allowed us to describe the molecular basis for enantioselectivity and to deduce the absolute configurations of all the four different stereoisomers (a–d) of finrozole. The ENA11His antibody fragment selectively binds the SR (a) enantiomer from the racemic mixture of a and d-enantiomers, thus allowing separation from the pharmacologically most active RS enantiomer (d). In particular, Asp95 and Asn35 of the H-chain in the ENA11 His antibody seem to provide this specificity through hydrogen bonding.

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