Back to results list
Please use this identifier to cite or link to this item:
|Title:||Interdomain flexibility and interfacial integrity of β-lactamase inhibitory protein (BLIP) modulate its binding to class A β-lactamases||Authors:||Huang, L
|Issue Date:||Aug-2021||Source:||Journal of biological chemistry, Aug. 2021, v. 297, no. 2, 100980||Abstract:||β-Lactamase inhibitory protein (BLIP) consists of a tandem repeat of αβ domains conjugated by an interdomain loop and can effectively bind and inactivate class A β-lactamases, which are responsible for resistance of bacteria to β-lactam antibiotics. The varied ability of BLIP to bind different β-lactamases and the structural determinants for significant enhancement of BLIP variants with a point mutation are poorly understood. Here, we investigated the conformational dynamics of BLIP upon binding to three clinically prevalent class A β-lactamases (TEM1, SHV1, and PC1) with dissociation constants between subnanomolar and micromolar. Hydrogen deuterium exchange mass spectrometry revealed that the flexibility of the interdomain region was significantly suppressed upon strong binding to TEM1, but was not significantly changed upon weak binding to SHV1 or PC1. E73M and K74G mutations in the interdomain region improved binding affinity toward SHV1 and PC1, respectively, showing significantly increased flexibility of the interdomain region compared to the wild-type and favorable conformational changes upon binding. In contrast, more rigidity of the interfacial loop 135-145 was observed in these BLIP mutants in both free and bound states. Consistently, molecular dynamics simulations of BLIP exhibited drastic changes in the flexibility of the loop 135-145 in all complexes. Our results indicated for the first time that higher flexibility of the interdomain linker, as well as more rigidity of the interfacial loop 135-145, could be desirable determinants for enhancing inhibition of BLIP to class A β-lactamases. Together, these findings provide unique insights into the design of enhanced inhibitors.||Keywords:||β-lactamases
β-lactamase inhibitory protein (BLIP)
Hydrogen deuterium exchange mass spectrometry
Molecular dynamics simulation
|Publisher:||American Society for Biochemistry and Molecular Biology||Journal:||Journal of biological chemistry||ISSN:||0021-9258||EISSN:||1083-351X||DOI:||10.1016/j.jbc.2021.100980||Rights:||© 2021 THE AUTHORS. Published by Elsevier Inc on behalf of American Society for Biochemistry and Molecular Biology. This is an open access article under the CCBY license (http://creativecommons.org/licenses/by/4.0/).
The following publication Huang, L., So, P. K., Chen, Y. W., Leung, Y. C., & Yao, Z. P. (2021). Interdomain flexibility and interfacial integrity of β-lactamase inhibitory protein (BLIP) modulate its binding to class A β-lactamases. Journal of Biological Chemistry, 297(2) is available at https://doi.org/10.1016/j.jbc.2021.100980
|Appears in Collections:||Journal/Magazine Article|
Show full item record
Files in This Item:
|1-s2.0-S0021925821007821-main.pdf||2.95 MB||Adobe PDF||View/Open|
Citations as of May 15, 2022
Citations as of May 15, 2022
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.