Isoelectric Point Research Articles

Buffer Screening of Protein Formulations Using a Coarse-Grained Protocol Based on Medicinal Chemistry Interactions.

In drug and vaccine development, the designed protein formulation should be highly stable against the temperature, pH, buffer, excipients, and other environmental settings. Similarly, in a sensing unit, one needs to know how strongly two biomolecules bind to guide the design of the biorecognition unit accordingly. Typically, the community performs a series of experiments to thoroughly examine the parameter space, the so-called design-of-experiment (DoE) method, to identify the optimal formulation conditions. Unfortunately, extensive physical testing entails high costs, repeatability issues, and a lack of in-depth knowledge of the underlying mechanisms that affect the final outcome. To address these challenges, we developed a physics-based simulation protocol for buffer screening of protein formulations. We are introducing a coarse-grained molecular simulation protocol that consists of six different interactions. The so-called medicinal chemistry interactions (electrostatics, hydrophobicity, hydrogen bonding propensity, disulfide bonding, and water-water) are based on the physical nature of the protein's amino acid and the partitioning/polarity of any other chemical constituent. The protocol is applied in immunoglobulin-based monoclonal antibodies. We have analyzed the protein behavior as a function of acidity (pH) to discover the isoelectric point by solving the Poisson-Boltzmann equation in a mesoscale grid. To identify the conditions under which the protein oligomerizes in a given buffer, pH, temperature, and ionic strength, we are performing dissipative particle dynamics (DPD) simulations. The protocol allows researchers to reach the high time/space scales required to study protein formulations in their full complexity. Combined with the disruptive protein folding artificial intelligence (AI) algorithms that have been recently developed, the protocol creates a powerful digital framework for cultivating advanced pharmaceutical and biological applications.

Microencapsulation of a green microalga (Chlorella vulgaris) by complex coacervation for its valuation as a feasible ingredient in pear snacks

The nutritional and bioactivity values of Chlorella vulgaris make this green microalga a potential functional food ingredient. However, its unpleasant taste and smell, as well as dark green color, have been considered restrictive for its consumers' acceptability. Here, it was hypothesized that complex coacervation can be used to produce microcapsules able to mitigate the sensory weaknesses of C. vulgaris in think pear snacks, while ensuring their acceptance and intention to buy. For this, two proteins with similar isoelectric points were used, the pea protein isolate (PPI) and the bovine serum albumin (BSA), while the partially-sulfated carrageenan (C) was the selected polysaccharide to encapsulate C. vulgaris (6 % w/w), at pH 3. Higher encapsulation yield (79 %) and efficiency (75 %) were obtained for PPI/C coacervates due to these polymers' higher electrostatic interaction at pH 3, contrasting with the higher repulsion promoted by some negatively charged BSA amino acids. Despite this, both microcapsules exhibited an amorphous structure and low water activity (aw around 0.3). However, PPI/C microcapsules had significantly lower moisture and water solubility (11 % and 57 %, respectively), as well as continuous shells. Moreover, PPI/C avoided the early release of C. vulgaris compounds at gastric conditions ensuring its later release in the intestinal tract. When incorporated into think pear snacks, the higher sensory scores and buying intention were obtained for snacks with PPI/C microcapsules containing C. vulgaris. Therefore, complex coacervation, particularly of PPI/C, allowed to mitigate non-valued microalga sensory attributes contributing for the future acceptance and consumption of algae-based food products.

Structure and Function of Fermentation-Derived Bovine Lactoferrin Produced from Komagataella phaffii

Bovine lactoferrin (bLf) confers significant functional benefits for human health, but low concentrations in milk and high cost of commercial production limit availability and thus product application. Precision fermentation offers a solution to increase availability of biosimilar recombinant bLf (rbLf) thereby opening new opportunities for this high-value ingredient. To comply with regulatory requirements, we aimed to establish that rbLf from <i>K. phaffii</i> is substantially similar to native bLf in structure and key functions. Intact mass analysis showed a molecular weight of 84 kDa for rbLf, comparable to 82-83 kDa of bLf. LC-MS <i>N</i>-linked glycan profiling revealed predominantly high-mannose-based glycans on rbLf, similar to ~50% of bLf glycans. The isoelectric point and core amino acid sequence of rbLf and bLf are identical. rbLf retains the functional ability to bind and release iron, bind to intestinal Lf receptors, increase epithelial cell growth (>120% of control, P < 0.0001), reduce EPEC growth (>50% reduction, P < 0.0001), bind LPS (+4 fold, P < 0.001) and antagonize LPS-induced TLR4 activity (>40% reduction, P < 0.0001). These results demonstrate similarity of rbLf in structure and function to native bLf, supporting the effective application for expanded market opportunities for infant and adult health.

Surface and electrochemical characteristics of S-scheme nanoheterostructured photocatalysts of AgIO3/Cu2SnS3 with enhanced solar energy driven photocatalytic activity

Wastewater treatment is regarded as one of the most challenges to overcome worldwide water threats. As a result, great efforts were devoted to find alterative solutions to maximize the usage of wastewater in industries or agriculture which consumes 70 % of water resources. Interestingly, the utilization of solar energy as a renewable, green and costless energy source is the ideal sustainable solution for wastewater treatment. In this context, Chalcogenides based nanoheterostructures such as AgIO3/Cu2SnS3 were prepared via simple, cost-effective and large-scale methods then utilized as visible light active photocatalysts for wastewater treatment. Scanning electron microscopy (SEM), X-ray powder diffraction (XRD), N2 sorpometry (BET), X-ray photoelectron spectroscopy (XPS), ultraviolet visible light spectrophotometry (UV–vis), and Isoelectric point (pHiep) were utilized to investigate the characteristics of nanoheterostructures. The produced nanoheterostructures were evaluated as photocatalysts for amoxicillin photodegradation using solar energy. The AgIO3/Cu2SnS3 (25ACS) demonstrated a superior photodegradation efficiency (93.5 %) after 60 min. Mot-Schottky plots and trapping experiments were carried out to have an extensive insight of the photodegradation mechanism.

Repairment of carbon steel oxide scale using Ce³⁺ and Ni²⁺ doped sol-gel technique

Corrosion of carbon steel rebars in concrete structures significantly compromises their safety, reliability, and environmental performance. This work focuses on enhancing rebar corrosion resistance by repairing the defects of oxide scale. Here, sol-gel was employed as a carrier, by which Ce3+ and Ni2+ were transferred to the oxide scale defects based on the isoelectric point theory, and deposited at these defects with the pH variation as temperature. The repaired oxide scale showed enhanced uniform elemental distribution and improved electrochemical properties, maintaining integrity even under prolonged exposure to Cl−-rich environments. Notably, the impedance modulus at 0.01 Hz (|Z | 0.01Hz) of treated samples was six times higher than untreated ones, indicating superior performance against high electric fields. This strategy shows great potential for enhancing the durability of concrete structures.

PH-induced interface protein structure changes to adjust the stability of tilapia protein isolate emulsion prepared by high-pressure homogenization

The pH is a crucial external factor affecting the structure and emulsification characteristics of proteins. The current study aimed to reveal the correlation between the secondary structure changes and tilapia protein isolate (TPI) emulsion stability under different pH (3.0–10.0) prepared by high-pressure homogenization. The results showed that TPI with significantly increased solubility and emulsifying properties when the pH keep away from the isoelectric point (pH 5.0). Meanwhile, TPI emulsions presented significantly enhanced stability (with decreased particle size, increased zeta potential, creaming index close to 0, and uniform dispersion of droplets) at pH 3.0 and 10.0. Interface-adsorbed protein mainly consists of a myosin-heavy chain and actin, and the secondary structure was significantly influenced by pH and high-pressure homogenization. The α-helix will be transformed into β-sheet and β-turn when pH is closer to pH 5.0. However, the high-pressure homogenization induced α-helix conversion to β-sheet. The correlation analysis revealed that emulsion stability is positively correlated with α-helix and negatively correlated with β-sheet. This work provides a deep insight into the correlation between secondary structure changes and the stability of TPI emulsion as affected by pH to offer an alternative way to enhance TPI emulsion stability.

Proteomic analysis of two novel peptides from the Odontobuthus doriae scorpion venom

The venom of the Odontobuthus doriae scorpion, prevalent in East Asia and Iran, has not been fully characterized. This study provides the first proteomic profile of O. doriae venom to explore its potential as a medical. 2D-PAGE analysis revealed 96 protein spots with isoelectric points from 3 to 9 and molecular weights between 6.6 to 205 kDa. Fourteen toxin fractions were isolated via HPLC, and SDS-PAGE showed seven protein bands ranging from 3.8 to 182 kDa. MALDI-TOF MS identified Peptide 1 and Peptide 2, resembling Hemoglobin beta-2 chain and Chaperonin HSP60 and suggest potential therapeutic applications for P1 and P2.

Characterization, expression and functional analysis of Hsp40 during LPS challenge in blood parrot Amphilophus citrinellus ×Vieja melanura

Heat shock protein 40 belonging to heat shock protein family plays an important role in the immune responses of organisms. In this study, the full length cDNA of Hsp40 was 2426 bp including a 1368 bp open reading frame (ORF) encoding 455 amino acids with a molecular weight of 49.16 kDa and a theoretical isoelectric point of 9.34 in blood parrot Vieja synspila ♀ × Amphilophus citrinellus ♂, an important ornamental fish in China. It had three conserved domains DnaJ, CRR and DnaJ_C. Phylogenetic analysis showed that the sequence of Hsp40 among species was conserved, and the blood parrot Hsp40 was closely related to Neolamprologus brichardi. Blood parrot Hsp40 mRNA could be detected in all of the tissues examined and mainly distributed in the cytoplasm. The expression of Hsp40 was upregulated during lipopolysaccharide (LPS) challenge. Upregulated Hsp40 inhibited the activity of nuclear factor κB (NF-κB) and activated protein 1 (AP-1) and reduced the ratio of Bax/Bcl-2 mRNA expression. This study provides a theoretical basis for further exploring the role of Hsp40 gene in the anti-bacterial immunity of blood parrot.

Key Control Characteristics of Carbon Black Materials for Fuel Cells and Batteries for a Standardized Characterization of Surface Properties

AbstractCarbon Blacks (CBs) are primarily used in electrocatalysis as support materials for nanoparticulate electrocatalysts. In fuel cells, CBs, catalysts, and proton conductive polymers (ionomers) undergo a dispersion/homogenization process for electrode coating. The interactions between components in dispersion and the surface of CB particles are complex. The surface characteristics of CB particles impact slurry formulation, electrode coating, and the resulting electrode layer, affecting fuel cell and battery performance and durability. This study investigates commercially available CBs such as Vulcan XC72R, Ketjen Black EC‐300J, Ketjen Black EC‐600JD, and Super C65, focusing on application‐relevant key control characteristics (KCCs). The KCCs include physicochemical features like electrical conductivity (electron transport through the catalyst layer, CL), oxygen‐containing surface groups (hydrophilicity/phobicity of the CL, catalyst functionalization), surface basicity/acidity, ionomer adsorption, and particle dispersibility (catalyst ink stability). KCCs are determined by particle bulk electrical conductivity, Boehm titration (BT), isoelectric point (IEP), and Hansen solubility parameters (HSP). HSP, understood as similarity parameters for particles, indicate how nanoparticle surfaces interact with liquids during dispersion. The proposed KCCs serve as a starting point for characterizing CBs and guiding product design to obtain meaningful structure‐property relationships, essential for a successful energy transition.

A time-resolved investigation at multiple-length scales of the structure of liquid foam stabilized by albumins from pea

HypothesisThe structural details of foams made with pea albumins are affected by the pH of the initial solution and followed heat treatment. ExperimentsAn in situ, time-resolved investigation of foams prepared with pea albumins was conducted using small-angle neutron scattering (SANS) in combination with imaging and conductance measurements. Solutions were tested at pH three pH values (3, 4.5, and 8) before and after heating (90 °C for 1 and 5 min). FindingsThe characteristic structures present in the foam from the nano to the meso-scale differed during drainage depending on solution pH. Foams obtained at pH 3, had the largest bubble radius and thinnest plateau border, as well as the highest extent of liquid drainage. At pH 4.5, close to the isoelectric point of the proteins, foams displayed similar bubbles’ behavior to those at pH 8, but with the largest film thickness. In this case, the proteins were extensively aggregated. Heating of the solutions prior to foaming did not significantly affect the foam aging regardless of pH. The quantification of specific surface areas and film thickness over time without sample disruption shows to be a powerful approach to designing foam structures.

Comparison of cat stone matrix and cat urine proteomes to human calcium oxalate stone matrix and urine proteomes.

Cat calcium oxalate monohydrate kidney stone matrix proteome showed great similarity to human calcium oxalate monohydrate stone matrix proteome, but inference of mechanistic similarity was limited by the absence of cat urine proteomic data. In this study, urine proteome distributions were measured by the same methods in 7 healthy cats for comparison to both the published human urine and cat calcium oxalate stone matrix proteomes to assess for similar enrichment patterns in both species. Furthermore, proteomic distributions were determined in cat struvite stone matrix to test for similarity to calcium oxalate monohydrate stone matrix and urine proteomes. Cat urine proteins demonstrated a similar distribution of abundance as a function of isoelectric points or net charge to human urine samples, and consequently the similarly altered patterns of protein distributions seen in calcium oxalate monohydrate stone matrix seen from both cat and human stones likely derives from the same preferential adsorption mechanism. Furthermore, the fact that protein abundance patterns seen in cat struvite stone matrix samples differ from both urine and calcium oxalate monohydrate stone matrix proteomes in systematic ways suggests that a combination of protein-protein and protein crystal interactions underly the formation of the crystal aggregates that comprise stones.

Cationic tannins as aggregation agents in the flocculation on hematite–silica system

ABSTRACT This study aims to evaluate the use of commercial tannin derivatives, namely, tannin 1 (CT1), tannin 2 (CT2), tannin 3 (CT3), and extract of tannin produced from angico bark, as aggregation agent in hematite–silica systems. Hematite and quartz were used in aggregation tests at dosages of 1, 10, and 100 mg/L at pH values of 6–9. The zeta potential and Jar tests were conducted in the best conditions of aggregation tests. The condensed tannin content of the commercial reagents is approximately similar, and it exceeds the content found in the extract obtained from angico bark. CT3 demonstrated superior percentages of sedimented material for both minerals at a dosage of 10 mg/L and pH 7 (97.12% and 95.68% for hematite and quartz, respectively). The aggregation agent produced from the angico bark displayed good sedimentation results for hematite (>90%); however, the results for quartz were unsatisfactory. The isoelectric point shifts to pH 8 when each mineral was conditioned with CT3. The jar test revealed higher sedimentation of the compositions for the lowest velocity gradient (60 rpm) with low turbidity values in the supernatant; however, the lowest turbidity (24.71 NTU) is observed in the sample with a higher percentage of hematite.

Sustainable development of fishery resources: Precipitation of protein from surimi rinsing wastewater by low-temperature plasma

This study aimed to determine the impact of low-temperature plasma (LTP) on the protein stability and composition in surimi rinsing wastewater (SRW). When SRW (300 mL) was treated with LTP at a power of 420 W and a flow rate of 1.1 L/min for 106 s, the protein precipitation was 76.04 %, the pH was close to the estimated value of the isoelectric point (pI). In comparison with the pI precipitation treatment, non-precipitated proteins in the SRW after LTP precipitation treatment showed significant changes in amino acids susceptible to oxidation but had minor changes in the hydrophobic amino acid content. LTP showed a markedly differentiated response to the different protein types in the SRW, increasing the relative amounts of several enzyme proteins in the non-precipitated protein. The combined effect of the active ingredients provided by LTP on protein conformation and hydrophobic interactions may be responsible for this ‘screening’ phenomenon.

Exploring the glycoprotein washing fluid-assisted cleanup for the restoration of oil-contaminated shorelines with environmental integrity

Spilled oil in ocean can spread to the shoreline and cause long-term impacts on the shoreline's ecological environment. Therefore, removing oil accumulated on shorelines is crucial. This study proposed an innovative ovalbumin (OVA) fluid-assisted method for the cleanup of oiled shoreline substrates. The oil removal efficiency of OVA fluids was systematically investigated. Higher concentrations of OVA fluids effectively enveloped and immobilized the oil, aiding in its separation from the sand surface. The increased temperature reduced the viscosity of emulsions, facilitating improved flow and oil removal. High salinity promoted the creation of oil particle aggregates molecules and facilitated the release of oil from the sand surface. The factorial analysis demonstrated that a high salt environment significantly enhances the combined impact of temperature and pH on oil removal performance. Different methods for the responsive separation of washing effluents were studied, and the most effective separation method was adjusting the pH of effluents to 4.54 (the isoelectric point of OVA). Separated precipitates exhibited good decomposition efficiency through thermal decomposition and biodegradation. OVA fluids boast advantages, such as low cost, easy recyclability, and non-toxicity, while ensuring high oil removal efficiency and making them a promising eco-friendly technique for the cleanup of oiled shorelines.

Plant-based marbled salami analogs: Emulsion-loaded microgels embedded within protein-polysaccharide hydrogel matrices

The inherent molecular differences between plant and animal proteins make it challenging to accurately simulate the optical and mechanical properties of real meat products using plant-derived ingredients. Soft matter physics approaches are therefore being employed to direct the assembly of plant proteins, polysaccharides, and lipids into meat-like structures. The main focus of the current study was to use soft matter physics approaches to create plant-based analogs of marbled meat products, such as salami, chorizo, or pepperoni. These products have whitish fatty regions embedded within reddish/brown lean regions. In this study, we mimicked the fatty regions in salami using emulsion-loaded alginate beads and the lean regions using composite hydrogels assembled from potato protein and gellan gum. Initially, the textural attributes of the plant-based lean regions were optimized by varying potato protein type (low or high isoelectric point) and concentration. As expected, the mechanical strength of the lean region analogs increased with increasing protein content and depended on protein type. Introducing alginate beads into the salami analogs had little impact on their textural attributes when measured under compression conditions but decreased their stiffness and fracture stress when measured under tensile conditions. In particular, introducing either small or large beads reduced the stiffness and breaking stress of the salami analogs, which was attributed to the introduction of weak points in the composite hydrogel. Finally, we showed that the appearance and textural attributes of real salami could be mimicked in plant-based salami by optimizing its composition and structure. This research provides valuable insights into methods of better simulating the properties of marbled meat products, like sausages and cold-cut meat products, thereby creating a more sustainable food supply.

Dependence of Protein Immobilization and Photocurrent Generation in PSI-FTO Electrodes on the Electrodeposition Parameters.

This study investigates the immobilization of cyanobacterial photosystem I (PSI) from Synechocystis sp. PCC 6803 onto fluorine-doped tin oxide (FTO) conducting glass plates to create photoelectrodes for biohybrid solar cells. The fabrication of these PSI-FTO photoelectrodes is based on two immobilization processes: rapid electrodeposition driven by an external electric field and slower adsorption during solvent evaporation, both influenced by gravitational sedimentation. Deposition and performance of photoelectrodes was investigated by UV-Vis absorption spectroscopy and photocurrent measurements. We investigated the efficiency of PSI immobilization under varying conditions, including solution pH, applied electric field intensity and duration, and electrode polarization, with the goals to control (1) the direction of migration and (2) the orientation of the PSI particles on the substrate surface. Variation in the pH value of the PSI solution alters the surface charge distribution, affecting the net charge and the electric dipole moment of these proteins. Results showed PSI migration to the positively charged electrode at pH 6, 7, and 8, and to the negatively charged electrode at pH 4.4 and 5, suggesting an isoelectric point of PSI between 5 and 6. At acidic pH, the electrophoretic migration was largely hindered by protein aggregation. Notably, photocurrent generation was consistently cathodic and correlated with PSI layer thickness, and no conclusions can be drawn on the orientation of the immobilized proteins. Overall, these findings suggest mediated electron transfer from FTO to PSI by the used electrolyte containing 10 mM sodium ascorbate and 200 μM dichlorophenolindophenol.

Exploring the Roles of the Swi2/Snf2 Gene Family in Maize Abiotic Stress Responses.

The maize Snf2 gene family plays a crucial role in chromatin remodeling and response to environmental stresses. In this study, we identified and analyzed 35 members of the maize Snf2 gene family (ZmCHR1 to ZmCHR35) using the Ensembl Plants database. Each protein contained conserved SNF2-N and Helicase-C domains. Phylogenetic analysis revealed six groups among the Snf2 proteins, with an uneven distribution across subfamilies. Physicochemical analysis indicated that the Snf2 proteins are hydrophilic, with varied amino acid lengths, isoelectric points, and molecular weights, and are predominantly localized in the nucleus. Chromosomal mapping showed that these genes are distributed across all ten maize chromosomes. Gene structure analysis revealed diverse exon-intron arrangements, while motif analysis identified 20 conserved motifs. Collinearity analysis highlighted gene duplication events, suggesting purifying selection. Cis-regulatory element analysis suggested involvement in abiotic and biotic stress responses. Expression analysis indicated tissue-specific expression patterns and differential expression under various stress conditions. Specifically, qRT-PCR validation under drought stress showed that certain Snf2 genes were upregulated at 12 h and downregulated at 24 h, revealing potential roles in drought tolerance. These findings provide a foundation for further exploration of the functional roles of the maize Snf2 gene family in development and stress responses.

Identification of subtelomeric cluster-genes associated to sexual stage in Toxoplasma gondii

Toxoplasma gondii is an obligate intracellular parasite with sexual reproduction in the intestinal epithelium of felines. The depletion of two gene repressors, AP2XI-2 and AP2XII-1, induces merozoite formation and gene expression towards sexual commitment. Based on RNA-seq datasets of AP2XI-2 and AP2XII-1 knock downs we identified subtelomeric (ST) TgB12 and hypothetical (HP) genes upregulated. Some of the differentially expressed genes (DEGs) are arranged in ST clusters. These DEG products are characterized by high isoelectric points (pI) and may encode small proteins. The potential roles of these clusters of DEG ST genes in environmental resistance or parasite sexual development of T. gondii is discussed.

Knockdown of the glucosamine-6-phosphate N-acetyltransferase gene by RNA interference enhances the virulence of entomopathogenic fungi against rice leaffolder Cnaphalocrocis medinalis

Insect cuticle acts as a first line of defense and a physical protective barrier against entomopathogens. Chitin biosynthesis pathway plays a crucial role in chitin formation in the cuticle of insects. Glucosamine-6-phosphate N-acetyltransferase (GNA) is a key enzyme in insect chitin biosynthesis that regulate the chitin formation. However, how GNA-mediated cuticle metabolism influences virulence of entomopathogenic fungi is still unknown. In this study, CmGNA gene was cloned and characterized from the rice leaffolder Cnaphalocrocis medinalis. The CmGNA contains an open read frame (ORF) 600 nucleotides, encoding 199 amino acids with an isoelectric point of 8.65 and a molecular weight of 22.30 kDa. The expression profile showed that CmGNA was highly expressed in 4th instar larvae and in the cuticle. Here, we also reported the impact of CmGNA gene and entomopathogenic fungi, Metarhizium anisopliae and Beauveria bassiana, on expression pattern of chitin biosynthesis genes, feeding behavior, survival rate and average body weight of infected larvae, phenotypic deformities, rate of pupation, and adult emergence. Our results showed that knockdown of CmGNA and application of M. anisopliae and B. bassiana three days after RNA interference (RNAi) significantly decreased the expression of CmGNA and other associated genes, reduced feeding efficiency and survival rate, and caused loss of average body weight, less rate of pupation and adult emergence of infected larvae. Knockdown of CmGNA gene also increased the lethality of larvae caused by M. anisopliae and B. bassiana and resulted in significantly phenotypic deformities of infected larvae. Our findings illustrated that RNAi-mediated CmGNA knockdown disturbed the chitin synthesis genes that led to enhancing the virulence of M. anisopliae and B. bassiana, which can provide us new insights to develop novel biocontrol strategies against C. medinalis.

Effect of PKD1L1 Mutation on its Interaction with PKD2 to Cause Situs Inversus Totalis

Situs inversus totalis (SIT) is a rare homozygous recessive disease caused by the mutation in PKD1L1, which is required for normal interaction with PKD2 and leads to different complications such as respiratory disorders, brain disorders and even obesity. The present study was designed to find out the mutational effect on the binding of PKD2 with mutated PKD1L1, which leads to SIT. The three-dimensional (3D) structure of wild type and mutated PKD1L1 was predicted with > 90% confidence using different online tools. The different online tools that were employed were SWISS-MODEL, Phyre2 (normal & intensive) and i-TASSER. To compute the physiochemical properties of PKD1L1 (wild & mutated) and PKD2 in silico approaches were employed using the ExPASy ProtParam tool. Physicochemical properties such as molecular weight, isoelectric point, the total number of negatively and positively charged residues, extinction coefficient, half-life, instability and aliphatic index, grand average of hydropathicity, and amino acid percentage were calculated. A lot of variability was observed in these parameters among PKD1L1 and PKD2, which accounted for diversification in their functional properties. The theoretical pI points showed that PKD1L1 (whole) is more basic with 6.64 pI compared to its first chain TOPO_DOM (amino acids from 1–1748) has a pI of 5.62 which means it is basic while PKD2 have the lowest pI point of 5.34. Docking was performed using the PatchDock and ClusPro online tools.