title: Role of κ→λ light-chain constant-domain switch in the structure and functionality of A17 reactibody.
abstract: The engineering of catalytic function in antibodies requires precise information on their structure. Here, results are presented that show how the antibody domain structure affects its functionality. The previously designed organophosphate-metabolizing reactibody A17 has been re-engineered by replacing its constant κ light chain by the λ chain (A17λ), and the X-ray structure of A17λ has been determined at 1.95 Å resolution. It was found that compared with A17κ the active centre of A17λ is displaced, stabilized and made more rigid owing to interdomain interactions involving the CDR loops from the VL and VH domains. These VL/VH domains also have lower mobility, as deduced from the atomic displacement parameters of the crystal structure. The antibody elbow angle is decreased to 126° compared with 138° in A17κ. These structural differences account for the subtle changes in catalytic efficiency and thermodynamic parameters determined with two organophosphate ligands, as well as in the affinity for peptide substrates selected from a combinatorial cyclic peptide library, between the A17κ and A17λ variants. The data presented will be of interest and relevance to researchers dealing with the design of antibodies with tailor-made functions.
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117871, Russian Federation.
authors.forename: Natalia
authors.initials: N
authors.lastname: Ponomarenko
authors.affiliation: European Molecular Biology Laboratory, Hamburg Unit, c/o DESY, Notkestrasse 85, 22603 Hamburg, Germany.
authors.forename: Spyros D
authors.initials: SD
authors.lastname: Chatziefthimiou
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117871, Russian Federation.
authors.forename: Inna
authors.initials: I
authors.lastname: Kurkova
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117871, Russian Federation.
authors.forename: Yuliana
authors.initials: Y
authors.lastname: Mokrushina
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117871, Russian Federation.
authors.forename: Yuliana
authors.initials: Y
authors.lastname: Mokrushina
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117871, Russian Federation.
authors.forename: Anastasiya
authors.initials: A
authors.lastname: Stepanova
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117871, Russian Federation.
authors.forename: Ivan
authors.initials: I
authors.lastname: Smirnov
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117871, Russian Federation.
authors.forename: Marat
authors.initials: M
authors.lastname: Avakyan
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117871, Russian Federation.
authors.forename: Tatyana
authors.initials: T
authors.lastname: Bobik
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117871, Russian Federation.
authors.forename: Azad
authors.initials: A
authors.lastname: Mamedov
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, Russian Federation.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Mitkevich
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117871, Russian Federation.
authors.forename: Alexey
authors.initials: A
authors.lastname: Belogurov
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Novosibirsk 630090, Russian Federation.
authors.forename: Olga S
authors.initials: OS
authors.lastname: Fedorova
authors.affiliation: St Petersburg Academic University, St Petersburg 194021, Russian Federation.
authors.forename: Michael
authors.initials: M
authors.lastname: Dubina
authors.affiliation: Lomonosov Moscow State University, Moscow 119991, Russian Federation.
authors.forename: Andrey
authors.initials: A
authors.lastname: Golovin
authors.affiliation: European Molecular Biology Laboratory, Hamburg Unit, c/o DESY, Notkestrasse 85, 22603 Hamburg, Germany.
authors.forename: Victor
authors.initials: V
authors.lastname: Lamzin
authors.affiliation: Université de Technologie de Compiègne, Unité Mixte de Recherche 6022, Centre National de la Recherche Scientifique, 60205 Compiègne, France.
authors.forename: Alain
authors.initials: A
authors.lastname: Friboulet
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, Russian Federation.
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Makarov
authors.affiliation: European Molecular Biology Laboratory, Hamburg Unit, c/o DESY, Notkestrasse 85, 22603 Hamburg, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Wilmanns
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117871, Russian Federation.
authors.forename: Alexander
authors.initials: A
authors.lastname: Gabibov
mesh_terms: D022801:Complementarity Determining Regions / Q000737:chemistry / Q000235:genetics; D003460:Crystallization; D018360:Crystallography, X-Ray; D006801:Humans; D007127:Immunoglobulin Constant Regions / Q000737:chemistry* / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000235:genetics; D007134:Immunoglobulin Switch Region / Q000235:genetics; D007145:Immunoglobulin kappa-Chains / Q000737:chemistry* / Q000235:genetics; D007146:Immunoglobulin lambda-Chains / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry; D013816:Thermodynamics
chemical_list: D022801:Complementarity Determining Regions; D007127:Immunoglobulin Constant Regions; D007143:Immunoglobulin Heavy Chains; D007145:Immunoglobulin kappa-Chains; D007146:Immunoglobulin lambda-Chains; D011994:Recombinant Proteins
keywords: A17 reactibody; antibodies