2WUR

Atomic resolution structure of GFP measured on a rotating anode

Summary for 2WUR

• Entry DOI: 10.2210/pdb2wur/pdb
• Related: 1B9C 1BFP 1C4F 1CV7 1EMA 1EMB 1EMC 1EME 1EMF 1EMG 1EMK 1EML 1EMM 1F09 1F0B 1GFL 1H6R 1HCJ 1HUY 1JBY 1JBZ 1JC0 1JC1 1KP5 1KYP 1KYR 1KYS 1MYW 1Q4A 1Q4B 1Q4C 1Q4D 1Q4E 1Q73 1QXT 1QY3 1QYF 1QYO 1QYQ 1RM9 1RMM 1RMO 1RMP 1RRX 1W7S 1W7T 1W7U 1YFP 1YHG 1YHH 1YHI 1YJ2 1YJF 1Z1P 1Z1Q 2AH8 2AHA 2B3P 2B3Q 2EMD 2EMN 2EMO 2FWQ 2FZU 2WSN 2WSO 2YFP
• Descriptor: GREEN FLUORESCENT PROTEIN, ISOPROPYL ALCOHOL, ETHANOL, ... (4 entities in total)
• Functional Keywords: chromophore, beta-barrel, luminescence, photoprotein, bioluminescence, fluorescent protein
• Biological source: AEQUOREA VICTORIA
• Total number of polymer chains: 1
• Total formula weight: 27252.78

Authors

Palm, G.J.,Schierbeek, A.J.,Kloos, M. (deposition date: 2009-10-07, release date: 2010-05-12, Last modification date: 2024-11-06)

Primary citation

Shinobu, A.,Palm, G.J.,Schierbeek, A.J.,Agmon, N.
Visualizing Proton Antenna in a High-Resolution Green Fluorescent Protein Structure.
J.Am.Chem.Soc., 132:11093-, 2010

PubMed Abstract: "Proton-collecting antenna" are conjectured to consist of several carboxylates within hydrogen-bond (HB) networks on the surface of proteins, which funnel protons to the orifice of an internal proton wire leading to the protein's active site. Yet such constructions were never directly visualized. Here we report an X-ray structure of green fluorescent protein (GFP) of the highest resolution to date (0.9 A). It allows the identification of some pivotal hydrogen atoms pertinent to uncertainties concerning the protonation state of the chromophore. Applying a computer algorithm for mapping proton wires in proteins reveals the previously observed "active site wire" connecting Glu222 with the surface carboxylate Glu5. In addition, it is now possible to identify what appears to be a proton-collecting apparatus of GFP. It consists of a negative surface patch containing carboxylates, threonines, and water molecules, connected by a HB network to Glu5. Furthermore, we detect exit points via Asn146 and His148 to a hydrophobic surface region. The more extensive HB network of the present structure, as compared with earlier GFP structures, is not accidental. A systematic investigation of over 100 mutants shows a clear correlation between the observed water content of GFP X-ray structures and their resolution. With increasing water content, the proton wires become progressively larger. These findings corroborate the scenario in which the photodissociated proton from wild-type GFP can leak outside, whereafter another proton is recruited via the proton-collecting apparatus reported herein.

PubMed: 20698675
DOI: 10.1021/JA1010652

Experimental method

X-RAY DIFFRACTION (0.9 Å)