Title: Structure and specificity of several triclocarban-binding single domain camelid antibody fragments

Abstract

The variable VHH domains of camelid single chain antibodies have been useful in numerous biotechnology applications due to their simplicity, biophysical properties, and abilities to bind to their cognate antigens with high affinities and specificity. Their interactions with proteins have been well-studied, but considerably less work has been done to characterize their ability to bind haptens. Here, a high-resolution structural study of three nanobodies (T4, T9, and T10) which have been shown to bind triclocarban (TCC, 3-(4-chlorophenyl)-1-(3,4-dichlorophenyl)urea) with near-nanomolar affinity shows that binding occurs in a tunnel largely formed by CDR1 rather than a surface or lateral binding mode seen in other nanobody-hapten interactions. Additional significant interactions are formed with a non-hypervariable loop, sometimes dubbed “CDR4”. A comparison of apo and holo forms of T9 and T10 shows that the binding site undergoes little conformational change upon binding of TCC. Structures of three nanobody-TCC complexes demonstrated there was not a standard binding mode. T4 and T9 have a high degree of sequence identity and bind the hapten in a nearly identical manner, while the more divergent T10 binds TCC in a slightly displaced orientation with the urea moiety rotated approximately 180° along the long axis of the molecule. In additionmore » to methotrexate, this is the second report of haptens binding in a tunnel formed by CDR1, suggesting that compounds with similar hydrophobicity and shape could be recognized by nanobodies in analogous fashion. Structure-guided mutations failed to improve binding affinity for T4 and T9 underscoring the high degree of natural optimization.« less

Authors:

Tabares-da Rosa, Sofia ^[1]; Wogulis, Linda A. ^[2]; Wogulis, Mark D. ^[2]; González-Sapienza, Gualberto  ^[1];

• Search DOE PAGES for author "González-Sapienza, Gualberto"
• Search DOE PAGES for ORCID "0000-0003-1824-0611"

Wilson, David K.  ^[2]

• Search DOE PAGES for author "Wilson, David K."
• Search DOE PAGES for ORCID "0000-0001-5997-1408"

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+ Show Author Affiliations

1. Univ. of the Republic (UDELAR), Montevideo (Uruguay)
2. Univ. of California, Davis, CA (United States)

Publication Date:

Mon Jul 23 00:00:00 EDT 2018

Research Org.:

Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS); SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States). Stanford Synchrotron Radiation Lightsource (SSRL); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States). Advanced Light Source (ALS)

Sponsoring Org.:

USDOE Office of Science (SC), Biological and Environmental Research (BER); USDOE Office of Science (SC), Basic Energy Sciences (BES); National Institutes of Health (NIH); Univ. of California; National Science Foundation (NSF)

OSTI Identifier:

1569914

Alternate Identifier(s):

OSTI ID: 1461214

Grant/Contract Number:  

AC02-06CH11357; AC02-76SF00515; AC02-05CH11231; P41GM103393; GM105404; GM073210; GM082250; GM094625; P41GM103403; S10OD021527; MR-15-328599; 1330685

Resource Type:

Accepted Manuscript

Journal Name:

JMR. Journal of Molecular Recognition

Additional Journal Information:

Journal Volume: 32; Journal Issue: 1; Journal ID: ISSN 0952-3499

Publisher:

Wiley

Country of Publication:

United States

Language:

ENGLISH

Subject:

59 BASIC BIOLOGICAL SCIENCES