scholarly journals Biophysical Characterization of Cancer-Related Carbonic Anhydrase IX

2020 ◽  
Vol 21 (15) ◽  
pp. 5277
Author(s):  
Katarina Koruza ◽  
A. Briana Murray ◽  
Brian P. Mahon ◽  
Jesse B. Hopkins ◽  
Wolfgang Knecht ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Carbonic Anhydrase ◽  
Small Angle ◽  
Catalytic Domain ◽  
Carbonic Anhydrase Ix ◽  
Biophysical Characterization ◽  
X Ray ◽  
Ca Ix ◽  
X Ray Scattering ◽  
Ray Scattering

Upregulation of carbonic anhydrase IX (CA IX) is associated with several aggressive forms of cancer and promotes metastasis. CA IX is normally constitutively expressed at low levels in selective tissues associated with the gastrointestinal tract, but is significantly upregulated upon hypoxia in cancer. CA IX is a multi-domain protein, consisting of a cytoplasmic region, a single-spanning transmembrane helix, an extracellular CA catalytic domain, and a proteoglycan-like (PG) domain. Considering the important role of CA IX in cancer progression and the presence of the unique PG domain, little information about the PG domain is known. Here, we report biophysical characterization studies to further our knowledge of CA IX. We report the 1.5 Å resolution crystal structure of the wild-type catalytic domain of CA IX as well as small angle X-ray scattering and mass spectrometry of the entire extracellular region. We used matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry to characterize the spontaneous degradation of the CA IX PG domain and confirm that it is only the CA IX catalytic domain that forms crystals. Small angle X-ray scattering analysis of the intact protein indicates that the PG domain is not randomly distributed and adopts a compact distribution of shapes in solution. The observed dynamics of the extracellular domain of CA IX could have physiological relevance, including observed cleavage and shedding of the PG domain.

Small-Angle X-ray Scattering and Mass Spectrometry Studies of γ-Irradiated C60in Cyclohexane

1996 ◽  
Vol 100 (13) ◽  
pp. 5426-5432 ◽  
Author(s):  
S. J. Henderson ◽  
R. L. Hettich ◽  
R. N. Compton ◽  
G. Bakale
Keyword(s):  
Mass Spectrometry ◽  
Small Angle ◽  
X Ray ◽  
X Ray Scattering ◽  
Ray Scattering

Chaperone-client interactions between Hsp21 and client proteins monitored in solution by small angle X-ray scattering and captured by crosslinking mass spectrometry

2017 ◽  
Vol 86 (1) ◽  
pp. 110-123 ◽  
Author(s):  
Gudrun Rutsdottir ◽  
Morten I. Rasmussen ◽  
Peter Hojrup ◽  
Katja Bernfur ◽  
Cecilia Emanuelsson ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Small Angle ◽  
X Ray ◽  
X Ray Scattering ◽  
Client Proteins ◽  
Ray Scattering

scholarly journals Small-Angle X-ray Scattering Models of APOBEC3B Catalytic Domain in a Complex with a Single-Stranded DNA Inhibitor

Viruses ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 290
Author(s):  
Fareeda M. Barzak ◽  
Timothy M. Ryan ◽  
Maksim V. Kvach ◽  
Harikrishnan M. Kurup ◽  
Hideki Aihara ◽  
...  
Keyword(s):  
Small Angle ◽  
Catalytic Domain ◽  
Adaptive Immune Response ◽  
Saxs Data ◽  
Single Stranded Dna ◽  
X Ray ◽  
Adaptive Immune ◽  
X Ray Scattering ◽  
Binding Substrate ◽  
Ray Scattering

In normal cells APOBEC3 (A3A-A3H) enzymes as part of the innate immune system deaminate cytosine to uracil on single-stranded DNA (ssDNA) to scramble DNA in order to give protection against a range of exogenous retroviruses, DNA-based parasites, and endogenous retroelements. However, some viruses and cancer cells use these enzymes, especially A3A and A3B, to escape the adaptive immune response and thereby lead to the evolution of drug resistance. We have synthesized first-in-class inhibitors featuring modified ssDNA. We present models based on small-angle X-ray scattering (SAXS) data that (1) confirm that the mode of binding of inhibitor to an active A3B C-terminal domain construct in the solution state is the same as the mode of binding substrate to inactive mutants of A3A and A3B revealed in X-ray crystal structures and (2) give insight into the disulfide-linked inactive dimer formed under the oxidizing conditions of purification.

Mg2+-Dependent Compaction and Folding of Yeast tRNAPheand the Catalytic Domain of theB. subtilisRNase P RNA Determined by Small-Angle X-ray Scattering†

Biochemistry ◽  
2000 ◽  
Vol 39 (36) ◽  
pp. 11107-11113 ◽  
Author(s):  
Xingwang Fang ◽  
Kenneth Littrell ◽  
Xiao-jing Yang ◽  
Stephen J. Henderson ◽  
Sonke Siefert ◽  
...  
Keyword(s):  
Small Angle ◽  
Catalytic Domain ◽  
X Ray ◽  
X Ray Scattering ◽  
Ray Scattering

Head-to-tail and side-by-side oligomerization of human carbonic anhydrase II: a small angle X-ray scattering study

2001 ◽  
Vol 28 (2) ◽  
pp. 143-150 ◽  
Author(s):  
Marcelo Ceolı́n ◽  
Umbra Sabina Colombo ◽  
Marı́a Cecilia Frate ◽  
Eugenia Clérico ◽  
Erica Antón ◽  
...  
Keyword(s):  
Carbonic Anhydrase ◽  
Small Angle ◽  
Carbonic Anhydrase Ii ◽  
X Ray ◽  
X Ray Scattering ◽  
Scattering Study ◽  
Human Carbonic Anhydrase ◽  
Ray Scattering

scholarly journals Structural comparison of protiated, H/D-exchanged and deuterated human carbonic anhydrase IX

2019 ◽  
Vol 75 (10) ◽  
pp. 895-903 ◽  
Author(s):  
K. Koruza ◽  
B. Lafumat ◽  
M. Nyblom ◽  
B. P. Mahon ◽  
W. Knecht ◽  
...  
Keyword(s):  
Carbonic Anhydrase ◽  
Crystal Structures ◽  
Active Site ◽  
Catalytic Domain ◽  
Crystal Form ◽  
Carbonic Anhydrase Ix ◽  
Structural Comparison ◽  
X Ray ◽  
Ca Ix ◽  
Human Carbonic Anhydrase

Human carbonic anhydrase IX (CA IX) expression is upregulated in hypoxic solid tumours, promoting cell survival and metastasis. This observation has made CA IX a target for the development of CA isoform-selective inhibitors. To enable structural studies of CA IX–inhibitor complexes using X-ray and neutron crystallography, a CA IX surface variant (CA IXSV; the catalytic domain with six surface amino-acid substitutions) has been developed that can be routinely crystallized. Here, the preparation of protiated (H/H), H/D-exchanged (H/D) and deuterated (D/D) CA IXSV for crystallographic studies and their structural comparison are described. Four CA IXSV X-ray crystal structures are compared: two H/H crystal forms, an H/D crystal form and a D/D crystal form. The overall active-site organization in each version is essentially the same, with only minor positional changes in active-site solvent, which may be owing to deuteration and/or resolution differences. Analysis of the crystal contacts and packing reveals different arrangements of CA IXSV compared with previous reports. To our knowledge, this is the first report comparing three different deuterium-labelled crystal structures of the same protein, marking an important step in validating the active-site structure of CA IXSV for neutron protein crystallography.

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