6B2N

Crystal structure of TEM-1 beta-lactamase mutant M182N

6B2N の概要

• エントリーDOI: 10.2210/pdb6b2n/pdb
• 分子名称: Beta-lactamase TEM, SULFATE ION (3 entities in total)
• 機能のキーワード: antibiotic resistance, beta-lactamase, hydrolase
• 由来する生物種: Escherichia coli
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 116083.85

構造登録者

Jimah, J.R.,Tolia, N.H. (登録日: 2017-09-20, 公開日: 2018-01-17, 最終更新日: 2024-10-16)

主引用文献

Zimmerman, M.I.,Hart, K.M.,Sibbald, C.A.,Frederick, T.E.,Jimah, J.R.,Knoverek, C.R.,Tolia, N.H.,Bowman, G.R.
Prediction of New Stabilizing Mutations Based on Mechanistic Insights from Markov State Models.
ACS Cent Sci, 3:1311-1321, 2017

PubMed Abstract: Protein stabilization is fundamental to enzyme function and evolution, yet understanding the determinants of a protein's stability remains a challenge. This is largely due to a shortage of atomically detailed models for the ensemble of relevant protein conformations and their relative populations. For example, the M182T substitution in TEM β-lactamase, an enzyme that confers antibiotic resistance to bacteria, is stabilizing but the precise mechanism remains unclear. Here, we employ Markov state models (MSMs) to uncover how M182T shifts the distribution of different structures that TEM adopts. We find that M182T stabilizes a helix that is a key component of a domain interface. We then predict the effects of other mutations, including a novel stabilizing mutation, and experimentally test our predictions using a combination of stability measurements, crystallography, NMR, and measurements of bacterial fitness. We expect our insights and methodology to provide a valuable foundation for protein design.

PubMed: 29296672
DOI: 10.1021/acscentsci.7b00465

実験手法

X-RAY DIFFRACTION (2 Å)