3GMW

Crystal Structure of Beta-Lactamse Inhibitory Protein-I (BLIP-I) in Complex with TEM-1 Beta-Lactamase

Summary for 3GMW

• Entry DOI: 10.2210/pdb3gmw/pdb
• Related: 3GMU 3GMV 3GMX 3GMY
• Descriptor: B-lactamase, Beta-lactamase inhibitory protein BLIP-I, PHOSPHATE ION, ... (4 entities in total)
• Functional Keywords: protein-protein complex, antibiotic resistance, hydrolase, plasmid, protein binding
• Biological source: Escherichia sp. Sflu5; More
• Total number of polymer chains: 4
• Total formula weight: 92287.39

Authors

Lim, D.C.,Gretes, M.,Strynadka, N.C.J. (deposition date: 2009-03-15, release date: 2009-03-31, Last modification date: 2024-11-27)

Primary citation

Gretes, M.,Lim, D.C.,de Castro, L.,Jensen, S.E.,Kang, S.G.,Lee, K.J.,Strynadka, N.C.
Insights into positive and negative requirements for protein-protein interactions by crystallographic analysis of the beta-lactamase inhibitory proteins BLIP, BLIP-I, and BLP.
J.Mol.Biol., 389:289-305, 2009

PubMed Abstract: Beta-lactamase inhibitory protein (BLIP) binds a variety of beta-lactamase enzymes with wide-ranging specificity. Its binding mechanism and interface interactions are a well-established model system for the characterization of protein-protein interactions. Published studies have examined the binding of BLIP to diverse target beta-lactamases (e.g., TEM-1, SME-1, and SHV-1). However, apart from point mutations of amino acid residues, variability on the inhibitor side of this enzyme-inhibitor interface has remained unexplored. Thus, we present crystal structures of two likely BLIP relatives: (1) BLIP-I (solved alone and in complex with TEM-1), which has beta-lactamase inhibitory activity very similar to that of BLIP; and (2) beta-lactamase-inhibitory-protein-like protein (BLP) (in two apo forms, including an ultra-high-resolution structure), which is unable to inhibit any tested beta-lactamase. Despite categorical differences in species of origin and function, BLIP-I and BLP share nearly identical backbone conformations, even at loop regions differing in BLIP. We describe interacting residues and provide a comparative structural analysis of the interactions formed at the interface of BLIP-I.TEM-1 versus those formed at the interface of BLIP.TEM-1. Along with initial attempts to functionally characterize BLP, we examine its amino acid residues that structurally correspond to BLIP/BLIP-I binding hotspots to explain its inability to bind and inhibit TEM-1. We conclude that the BLIP family fold is a robust and flexible scaffold that permits the formation of high-affinity protein-protein interactions while remaining highly selective. Comparison of the two naturally occurring, distinct binding interfaces built upon this scaffold (BLIP and BLIP-I) shows that there is substantial variation possible in the subnanomolar binding interaction with TEM-1. The corresponding (non-TEM-1-binding) BLP surface shows that numerous favorable backbone-backbone/backbone-side-chain interactions with a protein partner can be negated by the presence of a few, strongly unfavorable interactions, especially electrostatic repulsions.

PubMed: 19332077
DOI: 10.1016/j.jmb.2009.03.058

Experimental method

X-RAY DIFFRACTION (2.1 Å)