Crystallization and preliminary X-ray crystallographic investigations on a βγ-crystallin domain of absent in melanoma 1 (AIM1), a protein from<i>Homo sapiens</i>

Penmatsa, Aravind; Rajini, Bheemreddy; Sharma, Yogendra; Sankaranarayanan, Rajan

doi:10.1107/s1744309106005380

Cited by 4 publications

(3 citation statements)

References 19 publications

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“…17 The purified protein was crystallized using a 2 μl + 2 μl mixture of protein and a reservoir solution containing 1.3-1.4 M sodium citrate, 0.1 M Hepes, pH 7.3-7.5, with nonpolar solvents such as isopropanol (1-3%), DMSO (2%) and salts such as ammonium sulfate (0.06 M) and sodium chloride (0.4 M) as additives. 18 Different heavy-atom compounds such as thiomersal, cadmium chloride and sodium iodide were initially attempted by soaking crystals in high concentrations (10-50 mM) of these solutions and observing the crystals for morphological changes. Both time of soak and concentration of the soak were altered to get welldiffracting isomorphous derivatives.…”

Section: Domain Structurementioning

confidence: 99%

Exploring the Limits of Sequence and Structure in a Variant βγ-Crystallin Domain of the Protein Absent in Melanoma-1 (AIM1)

Penmatsa

Wistow

Sharma

et al. 2008

Journal of Molecular Biology

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Betagamma-crystallins belong to a superfamily of proteins in prokaryotes and eukaryotes that are based on duplications of a characteristic, highly conserved Greek key motif. Most members of the superfamily in vertebrates are structural proteins of the eye lens that contain four motifs arranged as two structural domains. Absent in melanoma 1 (AIM1), an unusual member of the superfamily whose expression is associated with suppression of malignancy in melanoma, contains 12 betagamma-crystallin motifs in six domains. Some of these motifs diverge considerably from the canonical motif sequence. AIM1g1, the first betagamma-crystallin domain of AIM1, is the most variant of betagamma-crystallin domains currently known. In order to understand the limits of sequence variation on the structure, we report the crystal structure of AIM1g1 at 1.9 A resolution. Despite having changes in key residues, the domain retains the overall betagamma-crystallin fold. The domain also contains an unusual extended surface loop that significantly alters the shape of the domain and its charge profile. This structure illustrates the resilience of the betagamma fold to considerable sequence changes and its remarkable ability to adapt for novel functions.

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Section: Domain Structurementioning

confidence: 99%

Exploring the Limits of Sequence and Structure in a Variant βγ-Crystallin Domain of the Protein Absent in Melanoma-1 (AIM1)

Penmatsa

Wistow

Sharma

et al. 2008

Journal of Molecular Biology

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“…For each organism, we selected every protein annotated in UniProt as a γ-crystallin (or as βAand βB-crystallin, respectively) and containing either two (for the single-domain βγ-crystallin from Ciona intestinalis) or four (all other organisms) complete Greek key domains. A small number of truncated crystallins were excluded, as were some homologs of absent-in-melanoma-1 (AIM-1), which has several βγ-crystallin domains but is not a lens protein [109].…”

Section: Evolutionary and Structural Analysis Of Chordate βγ-Crystallinsmentioning

confidence: 99%

The Functional Significance of High Cysteine Content in Eye Lens γ-Crystallins

Serebryany,

Martin,

Takahashi

2024

Biomolecules

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Cataract disease is strongly associated with progressively accumulating oxidative damage to the extremely long-lived crystallin proteins of the lens. Cysteine oxidation affects crystallin folding, interactions, and light-scattering aggregation especially strongly due to the formation of disulfide bridges. Minimizing crystallin aggregation is crucial for lifelong lens transparency, so one might expect the ubiquitous lens crystallin superfamilies (α and βγ) to contain little cysteine. Yet, the Cys content of γ-crystallins is well above the average for human proteins. We review literature relevant to this longstanding puzzle and take advantage of expanding genomic databases and improved machine learning tools for protein structure prediction to investigate it further. We observe remarkably low Cys conservation in the βγ-crystallin superfamily; however, in γ-crystallin, the spatial positioning of Cys residues is clearly fine-tuned by evolution. We propose that the requirements of long-term lens transparency and high lens optical power impose competing evolutionary pressures on lens βγ-crystallins, leading to distinct adaptations: high Cys content in γ-crystallins but low in βB-crystallins. Aquatic species need more powerful lenses than terrestrial ones, which explains the high methionine content of many fish γ- (and even β-) crystallins. Finally, we discuss synergies between sulfur-containing and aromatic residues in crystallins and suggest future experimental directions.

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“…Gelsolins are actin regulatory proteins [17]. The first crystal structure of AIM1, “AIM1-g1”, confirmed that the protein is a calcium ion-dependent protein that can compete with other microbial homologs to bind calcium [19,20]. The second and third motifs of AIM1 have rarely been investigated and their functions remain unclear.…”

Section: Introductionmentioning

confidence: 99%

Purification and Functional Characterization of the C-Terminal Domain of the β-Actin-Binding Protein AIM1 In Vitro

Cheng

et al. 2018

Molecules

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The protein absent in melanoma 1 (AIM1) is a member of the βγ-crystal lens superfamily that is associated with the development of multiple cancers. The binding of AIM1 to β-actin affects the migration and invasion of prostate cancer epithelial cells. The C-terminus of AIM1 is required for the β-actin interaction. However, the characteristics of AIM1 in vitro and the interaction mode between AIM1 and β-actin remain unknown. We describe novel methods to prepare pure recombinant AIM1 and identify possible binding modes between AIM1 and β-actin; we also obtain the crystal of the first two βγ-crystallin domains of AIM1 (g1g2) for future structural biology research. We first express and purify AIM1 after cloning the sequence into a modified pET-28a_psp expression vector. Next, we define the minimum unit formed by the βγ-crystallin domain repeats that bound to β-actin and perform its physiological function. Finally, we made the structural model of the AIM1 g1g2 that can be used to guide future biomedical investigations and prostate cancer research.

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Crystallization and preliminary X-ray crystallographic investigations on a βγ-crystallin domain of absent in melanoma 1 (AIM1), a protein fromHomo sapiens

Cited by 4 publications

References 19 publications

Exploring the Limits of Sequence and Structure in a Variant βγ-Crystallin Domain of the Protein Absent in Melanoma-1 (AIM1)

Exploring the Limits of Sequence and Structure in a Variant βγ-Crystallin Domain of the Protein Absent in Melanoma-1 (AIM1)

The Functional Significance of High Cysteine Content in Eye Lens γ-Crystallins

Purification and Functional Characterization of the C-Terminal Domain of the β-Actin-Binding Protein AIM1 In Vitro

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