Open Dataset XSF9A2LF
Cellobiose Dehydrogenase (pH range)
The interdomain electron transfer between the catalytic flavodehydrogenase domain and the electron transferring cytochrome domain of cellobiose dehydrogenase plays an essential role in biocatalysis, biosensors, and biofuel cells as well as for its natural function as an auxiliary enzyme of lytic polysaccharide monooxygenase. We investigated the mobility of the cytochrome and dehydrogenase domain of cellobiose dehydrogenase (CDH), which are hypothesized to limit interdomain electron transfer in solution by small angle X-ray scattering (SAXS). CDH from Myriococcum thermophilum was used to study the CDH mobility at different pH. We showed an increase of CDH flexibility at higher pH, indicating alterations of domain mobility.
Experimental descriptionThe sample used for the experiments represents MtCDH IIA: gene cdh, UniProt A9XK88, the CYT domain belongs to CAZy family AA8, and the DH domain to CAZy family AA3_1. SEC-SAXS data were collected at the SIBYLS beamline 12.3.1 at Advanced Light Source (LBNL Berkeley, California, USA) at 1.27 Å wavelength with a Pilatus 2M detector 2.1 m sample-to-detector distance, corresponding to a q range from 0.01 to 0.4 Å-1. The scattering vector is defined as q = 4π sinθ/λ, where 2θ is the scattering angle, and λ is the X-ray wavelength. For the pH-profile measurements a 50 mM phosphate-citrate buffer system was used. A size-exclusion chromatography was directly coupled with a SAXS flow cell and a multi-angle light scattering (SEC-SAXS-MALS). For analyzing the data and computing the pair distance distribution function the program SCÅTTER (https://bl1231.als.lbl.gov/scatter/) was used. The SAXS frames recorded before the protein elution was used for the background subtraction.
File descriptiongMtCDH.. are the files for the glycosylated (native) CDH. dgMtCDH.. are the files for the deglycosylated CDH. 35-75 are the pH values 3.5-7.5.
2023-02-28
Published2023-03-14
Data collection techniqueSEC-SAXS
Journal DOI
Advanced Light Source
BeamlineSIBYLS BL12.3.1
Wavelength1.27 Å
Sample to Detector Distance2.1 m
Bettina Motycka
University of Natural Resources and Life Sciences, Vienna (BOKU)
Austria
Collaborators
Project LeaderMichal Hammel
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Complete Set of SAS Data Files
The complete SAS dataset is downloadable as a zip file.
- MtCDH_pH-set.zip Download
Individual SAS Data Files (total 8)
These are individual SAS data files that may be raw or processed (merged, etc.). These may or not be included in a zip file containing a larger dataset.
- Wt_35.dat Download
- Wt_45.dat Download
- Wt_55.dat Download
- Wt_65.dat Download
- Wt_75.dat Download
- wt_dg_MI35.dat Download
- wt_dg_MI55.dat Download
- wt_dg_MI75.dat Download
Supplemental Data and Supporting Materials (total 0)
These data and materials may include X-ray crystal structure coordinates, multi-angle light scattering data, .etc. It may also include additional details or methods pertaining to the SAXS experiments, or the researcher's interpretations of the results.
Open SAXS data analysis sites in a new tab using these links
SAXS Similarity SAXS FrameSliceSample:
- Macromolecule 1: MtCDH
- Sample Full Name: Cellobiose dehydrogenase
- Sample Type: Protein
- Source Organism: Myriococcum thermophilum
- Source Organism NCBI Taxonomy ID: 2588717
- Expression System: Pichia pastoris X-33
- Expression NCBI Taxonomy ID: 4922
- Uniprot ID: A9XK88
- Sequence or Chemical Formula:
- QNNVPNTFTDPDSGITFNTWGLDEDSPQTQGGFTFGVALPSDALTTDASEFIGYLKCARNDESGWCGISLGGPMTNSLLITAWPHEDTVYTSLRFATGYAMPDVYEGDAEITQVSSSVNSTHFSLIFRCKNCLQWSHGGSSGGASTSGGVLVLGWVQAFDDPGNPTCPEQITLQQHDNGMGIWGAQLNTDAASPSYTDWAAQATKTVTGDCEGPTETSVVGVPVPTGVSFDYIVVGGGAGGIPAADKLSEAGKSVLLIEKGFASTANTGGTLGPEWLEGHDLTRFDVPGLCNQIWVDSKGIACEDTDQMAGCVLGGGTAVNAGLWFKPYSLDWDYLFPDGWKYNDVQPAINRALSRIPGTDAPSTDGKRYYQEGFEVLSKGLAAGGWTSVTANNAPDKKNRTFAHAPFMFAGGERNGPLGTYFQTAKKRNNFDVWLNTSVKRVIREGGHITGVEVEPFRDGGYEGIVPVTKVTGRVILSAGTFGSAKILLRSGIGPEDQLEVVAASEKDGPTMIGNSSWINLPVGYNLDDHLNTDTVISHPDVVFYDFYEAWDDPIESDKNSYLESRTGILAQAAPNIGPMFWEEIVGADGIVRQLQWTARVEGSLGAPNGHTMTMSQYLGRGATSRGRMTITPSLTTIVSDVPYLKDPNDKEAVIQGIINLQNALQNVANLTWLFPNSTITPREYVESMVVSPSNRRSNHWMGTNKLGTDDGRKGGSAVVDLDTRVYGTDNLFVIDASIFPGVPTTNPTSYIVVAAEHASSRILALPDLEPVPKYGQCGGREWTGSFVCADGSTCEYQNEWYSQCL