Mistletoe lectin I in complex with galactose and lactose reveals distinct sugar-binding properties

The structures of mistletoe lectin I (ML‐I) from Viscum album complexed with lactose and galactose have been determined at 2.3 Å resolution and refined to R factors of 20.9% (Rfree = 23.6%) and 20.9 (Rfree = 24.6%), respectively. ML‐I is a heterodimer and belongs to the class of ribosome‐inactivatin...

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Veröffentlicht in:Acta crystallographica. Section F, Structural biology and crystallization communications Structural biology and crystallization communications, 2005-01, Vol.61 (1), p.17-25
Hauptverfasser: Mikeska, Ruth, Wacker, Roland, Arni, Raghuvir, Singh, Tej P., Mikhailov, Albert, Gabdoulkhakov, Azat, Voelter, Wolfgang, Betzel, Christian
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container_title Acta crystallographica. Section F, Structural biology and crystallization communications
container_volume 61
creator Mikeska, Ruth
Wacker, Roland
Arni, Raghuvir
Singh, Tej P.
Mikhailov, Albert
Gabdoulkhakov, Azat
Voelter, Wolfgang
Betzel, Christian
description The structures of mistletoe lectin I (ML‐I) from Viscum album complexed with lactose and galactose have been determined at 2.3 Å resolution and refined to R factors of 20.9% (Rfree = 23.6%) and 20.9 (Rfree = 24.6%), respectively. ML‐I is a heterodimer and belongs to the class of ribosome‐inactivating proteins of type II, which consist of two chains. The A‐chain has rRNA N‐glycosidase activity and irreversibly inhibits eukaryotic ribosomes. The B‐chain is a lectin and preferentially binds to galactose‐terminated glycolipids and glycoproteins on cell membranes. Saccharide binding is performed by two binding sites in subdomains α1 and γ2 of the ML‐I B‐chain separated by ∼62 Å from each other. The favoured binding of galactose in subdomain α1 is achieved via hydrogen bonds connecting the 4‐hydroxyl and 3‐hydroxyl groups of the sugar moiety with the side chains of Asp23B, Gln36B and Lys41B and the main chain of 26B. The aromatic ring of Trp38B on top of the preferred binding pocket supports van der Waals packing of the apolar face of galactose and stabilizes the sugar–lectin complex. In the galactose‐binding site II of subdomain γ2, Tyr249B provides the hydrophobic stacking and the side chains of Asp235B, Gln238B and Asn256B are hydrogen‐bonding partners for galactose. In the case of the galactose‐binding site I, the 2‐hydroxyl group also stabilizes the sugar–protein complex, an interaction thus far rarely detected in galactose‐specific lectins. Finally, a potential third low‐affinity galactose‐binding site in subunit β1 was identified in the present ML‐I structures, in which a glycerol molecule from the cryoprotectant buffer has bound, mimicking the sugar compound.
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ML‐I is a heterodimer and belongs to the class of ribosome‐inactivating proteins of type II, which consist of two chains. The A‐chain has rRNA N‐glycosidase activity and irreversibly inhibits eukaryotic ribosomes. The B‐chain is a lectin and preferentially binds to galactose‐terminated glycolipids and glycoproteins on cell membranes. Saccharide binding is performed by two binding sites in subdomains α1 and γ2 of the ML‐I B‐chain separated by ∼62 Å from each other. The favoured binding of galactose in subdomain α1 is achieved via hydrogen bonds connecting the 4‐hydroxyl and 3‐hydroxyl groups of the sugar moiety with the side chains of Asp23B, Gln36B and Lys41B and the main chain of 26B. The aromatic ring of Trp38B on top of the preferred binding pocket supports van der Waals packing of the apolar face of galactose and stabilizes the sugar–lectin complex. 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Section F, Structural biology and crystallization communications</title><addtitle>Acta Cryst. F</addtitle><description>The structures of mistletoe lectin I (ML‐I) from Viscum album complexed with lactose and galactose have been determined at 2.3 Å resolution and refined to R factors of 20.9% (Rfree = 23.6%) and 20.9 (Rfree = 24.6%), respectively. ML‐I is a heterodimer and belongs to the class of ribosome‐inactivating proteins of type II, which consist of two chains. The A‐chain has rRNA N‐glycosidase activity and irreversibly inhibits eukaryotic ribosomes. The B‐chain is a lectin and preferentially binds to galactose‐terminated glycolipids and glycoproteins on cell membranes. Saccharide binding is performed by two binding sites in subdomains α1 and γ2 of the ML‐I B‐chain separated by ∼62 Å from each other. 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F</addtitle><date>2005-01</date><risdate>2005</risdate><volume>61</volume><issue>1</issue><spage>17</spage><epage>25</epage><pages>17-25</pages><issn>1744-3091</issn><eissn>1744-3091</eissn><abstract>The structures of mistletoe lectin I (ML‐I) from Viscum album complexed with lactose and galactose have been determined at 2.3 Å resolution and refined to R factors of 20.9% (Rfree = 23.6%) and 20.9 (Rfree = 24.6%), respectively. ML‐I is a heterodimer and belongs to the class of ribosome‐inactivating proteins of type II, which consist of two chains. The A‐chain has rRNA N‐glycosidase activity and irreversibly inhibits eukaryotic ribosomes. The B‐chain is a lectin and preferentially binds to galactose‐terminated glycolipids and glycoproteins on cell membranes. Saccharide binding is performed by two binding sites in subdomains α1 and γ2 of the ML‐I B‐chain separated by ∼62 Å from each other. The favoured binding of galactose in subdomain α1 is achieved via hydrogen bonds connecting the 4‐hydroxyl and 3‐hydroxyl groups of the sugar moiety with the side chains of Asp23B, Gln36B and Lys41B and the main chain of 26B. The aromatic ring of Trp38B on top of the preferred binding pocket supports van der Waals packing of the apolar face of galactose and stabilizes the sugar–lectin complex. In the galactose‐binding site II of subdomain γ2, Tyr249B provides the hydrophobic stacking and the side chains of Asp235B, Gln238B and Asn256B are hydrogen‐bonding partners for galactose. In the case of the galactose‐binding site I, the 2‐hydroxyl group also stabilizes the sugar–protein complex, an interaction thus far rarely detected in galactose‐specific lectins. Finally, a potential third low‐affinity galactose‐binding site in subunit β1 was identified in the present ML‐I structures, in which a glycerol molecule from the cryoprotectant buffer has bound, mimicking the sugar compound.</abstract><cop>5 Abbey Square, Chester, Cheshire CH1 2HU, England</cop><pub>Munksgaard International Publishers</pub><pmid>16508080</pmid><doi>10.1107/S1744309104031501</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record>
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subjects AFFINITY
Amino Acid Sequence
Binding Sites
BONDING
BUFFERS
CHAINS
CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY
FASTENING
Galactose - chemistry
GLYCEROL
HYDROGEN
INTERACTIONS
Lactose - chemistry
Mistletoe
mistletoe lectin I
Models, Molecular
Molecular Sequence Data
MOLECULES
Plant Preparations - chemistry
Plant Proteins - chemistry
POTENTIALS
Protein Structure Communications
Protein Structure, Secondary
R FACTORS
Ribosome Inactivating Proteins, Type 2
ribosome-inactivation proteins
RINGS
SACCHAROSE
Sequence Alignment
Sequence Homology, Amino Acid
STOWING
sugar-binding sites
Toxins, Biological - chemistry
title Mistletoe lectin I in complex with galactose and lactose reveals distinct sugar-binding properties
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