Research Information

The CFTR Protein Distribution Core offers high-quality, full-length recombinant cystic fibrosis transmembrane conductance regulator (CFTR) proteins expressed in and purified from mammalian cells. Wild-type, stabilized and CF-causing CFTR variants are available. Any interested CF investigator from within or outside of the UAB scientific community may order protein or request related Core services.

The CFTR Protein Production Core is independently funded by the Cystic Fibrosis Foundation to Dr. John Kappes, PI and Director of the Core, and Dr. Zhengrong Yang, co-PI, capitalizing on the CFTR protein expression and purification expertise of their laboratory to offer recombinant CFTR proteins and cell/membrane biomass.

Up to milligram quantities of analytically pure, properly folded, and biochemically active protein can be provided. For example, 3D structures have been determined by cryoEM [1], all exhibit a clear thermal unfolding transition [2],[3]), high ATPase activity (except for those containing ATPase incompetent mutations) [2],[3] and channel function [2-4]. These proteins are suitable for biochemical, biophysical and structural studies.

Core Function #1: Provide full-length recombinant CFTR protein

A number of CFTR proteins are available that have been optimized for expression and are well-characterized. These include wild-type and structurally stabilized variants, such as ΔF508, G551D, E1371Q and H1402S, each in a stabilized background. See Table 1 for additional information.

Core Function #2: Provide full-length recombinant CFTR protein of a specific design requested by investigator.

At the request of the investigator, the Core has the capabilities to construct novel proteins of specific interest.

Core Function #3: Provide whole cell or cell membranes expressing specific CFTR protein as requested by investigator.

Investigators requiring cells or cell membranes for their research may make such a request.

Overview of the Ordering Process

1. Please contact John Kappes at johnkappes@uabmc.edu and Zhengrong (Wendy) Yang at yangzw@uab.edu to discuss your specific needs and/or questions.

2. After determining what is needed, the requestor will be sent a CFTR Protein Production Order Form to complete.

3. Upon receipt of the Order Form, the requestor will be contacted by either Dr. Kappes or Yang to finalize the request and will be provided with an estimated delivery date.

4. A nominal fee will be charged for each batch of protein, membranes or cells, which may be paid through a purchase order.

Requesting investigators outside of UAB will be required to have a Material Transfer Agreement (MTA) in place guaranteeing responsible use of the supplied proteins. Other routine administrative stipulations for outside users will be discussed at the time of initial contact.

1. “The allosteric effects of the F508del mutation in human CFTR elucidated by cryo-EM enabled by protein engineering,” presented by John F. Hunt, Columbia University, at the 16th ECFS Basic Science Conference Pre-Conference Meeting entitled “CFTR: new insights on structure and function and implications for modulation,” in Dubrovnik, Croatia, 27 March 2019. Manuscript in preparation.
2. Yang, Z., et al., Structural stability of purified human CFTR is systematically improved by mutations in nucleotide binding domain 1. Biochim Biophys Acta, 2018. 1860(5): p. 1193-1204.
3. Hildebrandt, E., et al., Specific stabilization of CFTR by phosphatidylserine. Biochim Biophys Acta, 2017. 1859(2): p. 289-293.
4. Hildebrandt, E., et al., A stable human-cell system overexpressing cystic fibrosis transmembrane conductance regulator recombinant protein at the cell surface. Mol Biotechnol, 2015. 57(5): p. 391-405