Biophysical characterization and molecular phylogeny of human KIN protein

Philippsen, Gisele Strieder; Caruso, Ícaro Putinhon; Neto, Quirino Alves de Lima; Duarte, Francisco Ferreira; Rando, Fabiana dos Santos; Gerhardt, Edileusa C. M.; Fernandez, Maria Aparecida; Seixas, Flávio Augusto Vicente

doi:10.1007/s00249-019-01390-3

Cited by 5 publications

(5 citation statements)

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“…The increase in the percentage of β-sheet/β-strand may be related to the formation of β intermolecular structures that contribute to the formation and stabilization of aggregates. Such behavior was also observed for human DNA/RNA-binding KIN protein (Pattaro Júnior et al 2019) and for human serum albumin (1980) (WETZEL et al 1980).…”

Section: Influence Of Ph On the Characterization Of Secondary Structu...mentioning

confidence: 53%

Characterization of Secondary Structure and Thermal Stability by Biophysical Methods of the D-alanyl,D-alanine Ligase B Protein from Escherichia coli

Philippsen

Caruso

Sá

et al. 2022

PPL

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Background: Peptidoglycan (PG) is a key structural component of the bacterial cell wall and interruption of its biosynthesis is a validated target for antimicrobials. Of the enzymes involved in PG biosynthesis, D-alanyl,D-alanine ligase B (DdlB), is responsible for the condensation of two alanines, forming D-Ala-D-Ala, which is required for subsequent extracellular transpeptidase crosslinking of the mature peptidoglycan polymer. Objectives: We aimed the biophysical characterization of recombinant Escherichia coli DdlB (EcDdlB), regarding parameters of melting temperature (Tm), calorimetry and van’t Hoff enthalpy changes of denaturation ( and ), as well as characterization of elements of secondary structure at three different pHs. Methods: DdlB was overexpressed in E. coli BL21 and purified by affinity chromatography. Thermal stability and structural characteristics of the purified enzyme were analyzed by circular dichroism (CD), differential scanning calorimetry and fluorescence spectroscopy. Results: The stability of EcDdlB increased with proximity to its pI of 5.0, reaching the maximum at pH 5.4 with Tm and of 52.68 ºC and 484 kJ.mol-1, respectively. Deconvolutions of the CD spectra at 20 ºC showed a majority percentage of α-helix at pH 5.4 and 9.4, whereas for pH 7.4, an equal contribution of β-structures and α-helices was calculated. Thermal denaturation process of EcDdlB proved to be irreversible with an increase in β-structures that can contribute to the formation of protein aggregates. Conclutions: Such results will be useful for energy minimization of structural models aimed at virtual screening simulations, providing useful information in the search for drugs that inhibit peptidoglycan synthesis.

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Section: Influence Of Ph On the Characterization Of Secondary Structu...mentioning

confidence: 53%

Characterization of Secondary Structure and Thermal Stability by Biophysical Methods of the D-alanyl,D-alanine Ligase B Protein from Escherichia coli

Philippsen

Caruso

Sá

et al. 2022

PPL

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“…1) of KIN17 was respectively 90.37 and 65.36%, which means that the two domains are highly conserved, whereas this conservative phenomenon in the KOW motif was only found in the higher eukaryote, speculating that the KOW motif appeared late in evolution. >50% of the secondary structure of KIN17 consists of random coil and β-turns, while the components of α-helices and β-strands make up for <50% of the structure of KIN17 (9). Function of KIN17.…”

Section: Kin17 a Dna And Rna Binding Proteinmentioning

confidence: 99%

Roles and regulatory mechanisms of KIN17 in cancers (Review)

Huang

Dai

et al. 2023

Oncol Lett

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KIN17, which is known as a DNA and RNA binding protein, is highly expressed in numerous types of human cancers and was discovered to participate in several vital cell behaviors, including DNA replication, damage repair, regulation of cell cycle and RNA processing. Furthermore, KIN17 is associated with cancer cell proliferation, migration, invasion and cell cycle regulation by regulating pathways including the p38 MAPK, NF-κB-Snail and TGF-β/Smad2 signaling pathways. In addition, knockdown of KIN17 was found to enhance the sensitivity of tumor cells to chemotherapeutic agents. Immunohistochemical analysis revealed that there were significant differences in the expression of KIN17 between cancer tissues and adjacent tissues. Both the Kaplan-Meier survival analysis and multivariate Cox regression analysis indicated that KIN17 is aberrantly high expressed in various tumor tissues and is also associated with poor prognosis in patients with various tumor types. Taken together, KIN17 has key roles in tumorigenesis and cancer development. Investigating the relationship between KIN17 and neoplasms will provide a vital theoretical basis for KIN17 to serve as a diagnostic and prognostic biomarker for cancer patients and as a potential target for cancer therapy.

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Section: Introductionmentioning

confidence: 99%

“…Biol. 2021, 1, FOR PEER REVIEW 2 The DNA-and RNA-binding protein KIN, also known as Kin17, is well conserved among species from lower to higher eukaryotes, suggesting an important role in the maintenance of basic cellular function which remains to be defined [9]. It was identified by antibodies raised against bacterial Escherichia coli repair protein RecA, in an effort to identify mammalian orthologs [10].…”

Section: Introductionmentioning

confidence: 99%

“…A recent biophysical evaluation revealed new insights into the recombinant human KIN protein, (HSAKIN), where the secondary structure is composed of more than 50% unfolded elements such as random coils and β-turns [9], suggesting a protein with high flexibility. Additionally, phylogenetic studies of the conserved sequences show that the KOW motif present in the KIN protein is conserved only in higher eukaryotes [9]. In the current manuscript, we present interactors of KIN identified by BioID, a proximity-dependent biotin identification methodology [23], in which a mutated biotin ligase (BirA*) is fused to our protein of interest, allowing for the local activation of biotin and subsequent biotinylation of proteins in its vicinity.…”

Section: Introductionmentioning

confidence: 99%

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Interactome Analysis of KIN (Kin17) Shows New Functions of This Protein

Gaspar

Ramos

Cloutier

et al. 2021

CIMB

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KIN (Kin17) protein is overexpressed in a number of cancerous cell lines, and is therefore considered a possible cancer biomarker. It is a well-conserved protein across eukaryotes and is ubiquitously expressed in all cell types studied, suggesting an important role in the maintenance of basic cellular function which is yet to be well determined. Early studies on KIN suggested that this nuclear protein plays a role in cellular mechanisms such as DNA replication and/or repair; however, its association with chromatin depends on its methylation state. In order to provide a better understanding of the cellular role of this protein, we investigated its interactome by proximity-dependent biotin identification coupled to mass spectrometry (BioID-MS), used for identification of protein–protein interactions. Our analyses detected interaction with a novel set of proteins and reinforced previous observations linking KIN to factors involved in RNA processing, notably pre-mRNA splicing and ribosome biogenesis. However, little evidence supports that this protein is directly coupled to DNA replication and/or repair processes, as previously suggested. Furthermore, a novel interaction was observed with PRMT7 (protein arginine methyltransferase 7) and we demonstrated that KIN is modified by this enzyme. This interactome analysis indicates that KIN is associated with several cell metabolism functions, and shows for the first time an association with ribosome biogenesis, suggesting that KIN is likely a moonlight protein.

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Biophysical characterization and molecular phylogeny of human KIN protein

Cited by 5 publications

References 60 publications

Characterization of Secondary Structure and Thermal Stability by Biophysical Methods of the D-alanyl,D-alanine Ligase B Protein from Escherichia coli

Characterization of Secondary Structure and Thermal Stability by Biophysical Methods of the D-alanyl,D-alanine Ligase B Protein from Escherichia coli

Roles and regulatory mechanisms of KIN17 in cancers (Review)

Interactome Analysis of KIN (Kin17) Shows New Functions of This Protein

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