Antiferritin VL homodimer binds human spleen ferritin with high specificity

Yvonne Nymalm, Zinaida Kravchuk, Tiina Salminen, Alexander A. Chumanevich, Anatoly P. Dubnovitsky, Jussi Kankare, Olli Pentikäinen, Jukka Lehtonen, Paolo Arosio, Sergey Martsev, Mark S. Johnson

    Research output: Contribution to journalArticleScientificpeer-review

    8 Citations (Scopus)

    Abstract

    The antiferritin variable light domain (VL) dimer binds human spleen ferritin ( approximately 85% L subunits) but with approximately 50-fold lower affinity, K(a)=4 x 10(7) x M(-1), than the parent F11 antibody (K(a)=2.1 x 10(9) x M(-1)). The VL dimer does not recognize either rL (100% L subunits) or rH (100% H subunits) human ferritin, whereas the parent antibody recognizes rL-ferritin. To help explain the differences in ferritin binding affinities and specificities, the crystal structure of the VL domain (2.8A resolution) was determined by molecular replacement and models of the antiferritin VL-VH dimer were made on the basis of antilysozyme antibody D1.3. The domain interface is smaller in the VL dimer but a larger number of interdomain hydrogen bonds may prevent rearrangement on antigen binding. The antigen binding surface of the VL dimer is flatter, lacking a negatively charged pocket found in the VL-VH models, contributed by the CDR3 loop of the VH domain. Loop CDR2 (VL dimer) is located away from the antigen binding site, while the corresponding loop of the VH domain would be located within the antigen binding site. Together these differences lead to 50-fold lower binding affinity in the VL dimer and to more restricted specificity than is seen for the parent antibody.

    Original languageEnglish
    Pages (from-to)171-86
    Number of pages16
    JournalJournal of Structural Biology
    Volume138
    Issue number3
    DOIs
    Publication statusPublished - Jun 2002
    MoE publication typeA1 Journal article-refereed

    Keywords

    • Amino Acid Sequence
    • Antibodies/chemistry
    • Binding Sites
    • Crystallography, X-Ray
    • Dimerization
    • Dose-Response Relationship, Drug
    • Ferritins/chemistry
    • Humans
    • Hydrogen Bonding
    • Kinetics
    • Models, Molecular
    • Molecular Sequence Data
    • Protein Binding
    • Protein Structure, Secondary
    • Protein Structure, Tertiary
    • Sequence Homology, Amino Acid
    • Spleen/metabolism

    Fingerprint

    Dive into the research topics of 'Antiferritin VL homodimer binds human spleen ferritin with high specificity'. Together they form a unique fingerprint.

    Cite this