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rabbit polyclonal prps1 (Proteintech)

Name: rabbit polyclonal prps1
Brand: Proteintech
Rating: 93 (15 reviews)

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Proteintech rabbit polyclonal prps1
Rabbit Polyclonal Prps1, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 93 stars, based on 15 article reviews

rabbit polyclonal prps1 - by Bioz Stars, 2026-02

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A Phylogenetic distribution of PRPS homologs in eukaryotes, with presence/absence of NHR-containing PRPS homologs in opisthokonts. PRPSAP2 denotes orthologs of mammalian PRPSAP2; Prs1 and Prs5 represent S. cerevisiae Prs1 and Prs5, respectively. Black asterisk indicates Class II presence across most metazoans, excluding Craniata. Red asterisks indicate Class II PRPS identified in only two Nuclearia species and one Chytridomycota species. B Conserved splice site junctions among PRPS homologs across different representative organisms in Opisthokonta. Gene structures for PRPS, PRPSAP2, and Prs5-encoding genes shown with exons as colored boxes; introns not displayed. Multiple sequence alignment (MSA) of translated sequences via Clustal Omega highlights a conserved splice site junction (red arrow) with adjacent amino acids shown. Top bar shows full H. sapiens PRPS1 with RF loop (red box) and CF loop (black box). Dotted red and black lines project corresponding RF and CF loops positions, respectively onto other homologs. Insertions in RF and CF loops of PRPSAP2 and Prs5 shown with red and black hatch marks, respectively (hatch marks not to scale; NHRs vary in length). C Phylogenetic distribution of PRPS homologs in chordates. Mostly present – indicates present in most taxa. Black asterisk highlights that Class II PRPS is found in most urochordates, except Oikopleura , which has additional Class I PRPS homologs not observed in other organisms from this clade. D , E Conserved splice site junctions between PRPS1 and PRPS2 ( D ), and PRPSAP2 and PRPSAP1 ( E ) across different representative organisms in jawed Vertebrata. Gene structures for PRPS, PRPSAP2, and Prs5-encoding genes shown with exons as colored boxes; introns not displayed. Dotted red and black lines denote RF and CF loop regions, respectively; insertions in PRPSAP2 and PRPSAP1 shown as red and black hatch marks, respectively (hatch marks not to scale; NHRs vary in length).

Journal: Nature Communications

Article Title: The role of gene duplication and paralog specialisation in the evolution of the mammalian PRPS complex

doi: 10.1038/s41467-025-61216-z

Figure Lengend Snippet: A Phylogenetic distribution of PRPS homologs in eukaryotes, with presence/absence of NHR-containing PRPS homologs in opisthokonts. PRPSAP2 denotes orthologs of mammalian PRPSAP2; Prs1 and Prs5 represent S. cerevisiae Prs1 and Prs5, respectively. Black asterisk indicates Class II presence across most metazoans, excluding Craniata. Red asterisks indicate Class II PRPS identified in only two Nuclearia species and one Chytridomycota species. B Conserved splice site junctions among PRPS homologs across different representative organisms in Opisthokonta. Gene structures for PRPS, PRPSAP2, and Prs5-encoding genes shown with exons as colored boxes; introns not displayed. Multiple sequence alignment (MSA) of translated sequences via Clustal Omega highlights a conserved splice site junction (red arrow) with adjacent amino acids shown. Top bar shows full H. sapiens PRPS1 with RF loop (red box) and CF loop (black box). Dotted red and black lines project corresponding RF and CF loops positions, respectively onto other homologs. Insertions in RF and CF loops of PRPSAP2 and Prs5 shown with red and black hatch marks, respectively (hatch marks not to scale; NHRs vary in length). C Phylogenetic distribution of PRPS homologs in chordates. Mostly present – indicates present in most taxa. Black asterisk highlights that Class II PRPS is found in most urochordates, except Oikopleura , which has additional Class I PRPS homologs not observed in other organisms from this clade. D , E Conserved splice site junctions between PRPS1 and PRPS2 ( D ), and PRPSAP2 and PRPSAP1 ( E ) across different representative organisms in jawed Vertebrata. Gene structures for PRPS, PRPSAP2, and Prs5-encoding genes shown with exons as colored boxes; introns not displayed. Dotted red and black lines denote RF and CF loop regions, respectively; insertions in PRPSAP2 and PRPSAP1 shown as red and black hatch marks, respectively (hatch marks not to scale; NHRs vary in length).

Article Snippet: After blocking, the membranes were washed and incubated overnight at 4 °C with primary antibodies (diluted 1:1000) prepared in 3% BSA in TBS-T. Primary antibodies used were: CAD (Cell Signaling #93925), TCP1-η (Santa Cruz #sc-271951), FASN (Cell Signaling #3180), FLC (Santa Cruz #sc-390558), HK2 (Cell Signaling #2867), AK2 (Santa Cruz #sc-374095), PRPS1/2 (Santa Cruz #sc-100822), PRPS1 (Proteintech #15549-1-AP), PRPS2 (Sigma #SAB2107995), PRPS1/2/3 (Santa Cruz #sc-376440), PRPSAP1 (Santa Cruz #sc-398422), PRPSAP2 (Proteintech #17814-1-AP), HSP90 (Cell Signaling #4877), β-Actin (Cell Signaling #4970; Cell Signaling #3700), ALFA-HRP (SynapticSystems # N1505-HRP), XO (Abcam #109235), Ras (G12V Mutant Specific) (Cell Signaling #14412), Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cell Signaling #4376), p44/42 MAPK (Erk1/2) (Cell Signaling #9102), β-Tubulin (Cell Signaling #2128), Phospho-AMPKα (Thr172) (Cell Signaling #2535), AMPKα (Cell Signaling #2532), cleaved PARP1 (Abcam #32064), GAPDH (Cell Signaling #5174), HPRT (Abcam #109021).

Techniques: Sequencing

A Structure of dimeric human PRPS1 (PDB: 2HCR), with a zoom-in of catalytic site highlighting metal binding site (Cd 2+ ), AMP (represents AMP moiety of ATP), SO 4 2− (represents 5′-phosphate of R5P), and several conserved active site residues (magenta). D171 coordinates metal binding, K194 interacts with ATP, R196, and T225 interact with R5P, and N200 stabilizes the catalytic loop. B WebLogo depicting MSA of active and regulatory site residues from PRPS1 and PRPSAP2 from representative organisms in opisthokonts ( n = 44 each). Numbers below indicate corresponding residue positions in human PRPS1 ( NP_002755.1 ) and PRPSAP2 ( NP_001340030.1 ). Asterisks denote residues conserved in PRPS but substituted in PRPSAP2 (also shown in A ). C AlphaFold2-predicted structure of human PRPSAP2 ( NP_001340030.1 ), with a zoom-in highlighting four non-conserved residues (magenta) at corresponding active site positions in PRPS1 shown in ( A ). AMP modeled to indicate putative ATP binding site. D Trimeric structure of human PRPS1 (PDB: 2HCR). Red and magenta residues in dashed box represent dimer interface residues in bent ( B , C ) and parallel ( A , B ) dimers, respectively. Amino acid sequence of B. subtilis PRPS aligned with Opisthokonta PRPS homologs to identify corresponding dimer interface residues for WebLogo. Representative sequences include PRPS1 ( n = 44), PRPSAP2 ( n = 44) from opisthokonts, and PRPS2 ( n = 46), PRPSAP1 ( n = 92) from jawed vertebrates. Residue numbers based on human PRPS1 ( NP_002755.1 ), PRPS2 ( NP_002756.1 ), PRPSAP1 ( AAH09012.1 ), and PRPSAP2 ( NP_001340030.1 ). E PRPS paralogs from S. cerevisiae , H. sapiens, R. potamoides, T. trahens, and D. rotans showing relative NHR positions in paralogs with expanded CF and/or RF loops. Open bar for each representative species represents full polypeptide sequences of their ancestral PRPS (bold). Amino acid positions corresponding to RF and CF loops labeled within bars. NHR insertions sites for Prs1, Prs5, PRPSAP1, PRPSAP2, PrsB, and PRPSAP-like homologs marked by triangles above/below bars. Numbers next to triangles indicate amino acid count per NHR.

Journal: Nature Communications

Article Title: The role of gene duplication and paralog specialisation in the evolution of the mammalian PRPS complex

doi: 10.1038/s41467-025-61216-z

Figure Lengend Snippet: A Structure of dimeric human PRPS1 (PDB: 2HCR), with a zoom-in of catalytic site highlighting metal binding site (Cd 2+ ), AMP (represents AMP moiety of ATP), SO 4 2− (represents 5′-phosphate of R5P), and several conserved active site residues (magenta). D171 coordinates metal binding, K194 interacts with ATP, R196, and T225 interact with R5P, and N200 stabilizes the catalytic loop. B WebLogo depicting MSA of active and regulatory site residues from PRPS1 and PRPSAP2 from representative organisms in opisthokonts ( n = 44 each). Numbers below indicate corresponding residue positions in human PRPS1 ( NP_002755.1 ) and PRPSAP2 ( NP_001340030.1 ). Asterisks denote residues conserved in PRPS but substituted in PRPSAP2 (also shown in A ). C AlphaFold2-predicted structure of human PRPSAP2 ( NP_001340030.1 ), with a zoom-in highlighting four non-conserved residues (magenta) at corresponding active site positions in PRPS1 shown in ( A ). AMP modeled to indicate putative ATP binding site. D Trimeric structure of human PRPS1 (PDB: 2HCR). Red and magenta residues in dashed box represent dimer interface residues in bent ( B , C ) and parallel ( A , B ) dimers, respectively. Amino acid sequence of B. subtilis PRPS aligned with Opisthokonta PRPS homologs to identify corresponding dimer interface residues for WebLogo. Representative sequences include PRPS1 ( n = 44), PRPSAP2 ( n = 44) from opisthokonts, and PRPS2 ( n = 46), PRPSAP1 ( n = 92) from jawed vertebrates. Residue numbers based on human PRPS1 ( NP_002755.1 ), PRPS2 ( NP_002756.1 ), PRPSAP1 ( AAH09012.1 ), and PRPSAP2 ( NP_001340030.1 ). E PRPS paralogs from S. cerevisiae , H. sapiens, R. potamoides, T. trahens, and D. rotans showing relative NHR positions in paralogs with expanded CF and/or RF loops. Open bar for each representative species represents full polypeptide sequences of their ancestral PRPS (bold). Amino acid positions corresponding to RF and CF loops labeled within bars. NHR insertions sites for Prs1, Prs5, PRPSAP1, PRPSAP2, PrsB, and PRPSAP-like homologs marked by triangles above/below bars. Numbers next to triangles indicate amino acid count per NHR.

Techniques: Binding Assay, Residue, Sequencing, Labeling

$A SDS-PAGE followed by Coomassie stain of eluates from GFP immunoprecipitation (IP) performed in NIH3T3 and HEK293T cells stably expressing PRPS1-GFP, PRPS2-GFP, and NES-GFP. NES (Nuclear export signal)-GFP used as control. Asterisk indicates a non-specific band in eluates. B Scatter plot from mass spectrometry (MS) runs of eluates from GFP IP in NIH3T3 ( x -axis) and HEK293T cells ( y -axis) stably expressing PRPS1-GFP. Axes represent square root-transformed SEQUEST HT scores of PRPS1-GFP normalized to control. PRPS1L1 isoform detected only in HEK29T cells. C Western blot analysis of size exclusion chromatography (SEC) fractions collected from NIH3T3 native whole cell lysates. Cell lysates were fractionated using a Superose 6 Increase 3.2/300 column. Immunoblots probing indicated PRPS complex members and internal standards shown. D–F Western blot analysis of SEC fractions from native tissue lysates of liver ( D ), kidney ( E ), and lung ( F ). Twelve-week-old male C57BL/6 mice were used. Lysates were fractionated on a Superose 6 Increase 3.2/300 column. Immunoblots probing PRPS complex members and internal standards are shown. Circular pictograms below SEC immunoblots schematize PRPS complex configurations; double circle denotes multiple copies of the protein interacting within the heteromeric complex. Coomassie staining ( A ) and western blot data ( C – F ) are representative of at least 3 biological repeats. Source data are provided as a Source Data file.$

Journal: Nature Communications

Article Title: The role of gene duplication and paralog specialisation in the evolution of the mammalian PRPS complex

doi: 10.1038/s41467-025-61216-z

Figure Lengend Snippet: A SDS-PAGE followed by Coomassie stain of eluates from GFP immunoprecipitation (IP) performed in NIH3T3 and HEK293T cells stably expressing PRPS1-GFP, PRPS2-GFP, and NES-GFP. NES (Nuclear export signal)-GFP used as control. Asterisk indicates a non-specific band in eluates. B Scatter plot from mass spectrometry (MS) runs of eluates from GFP IP in NIH3T3 ( x -axis) and HEK293T cells ( y -axis) stably expressing PRPS1-GFP. Axes represent square root-transformed SEQUEST HT scores of PRPS1-GFP normalized to control. PRPS1L1 isoform detected only in HEK29T cells. C Western blot analysis of size exclusion chromatography (SEC) fractions collected from NIH3T3 native whole cell lysates. Cell lysates were fractionated using a Superose 6 Increase 3.2/300 column. Immunoblots probing indicated PRPS complex members and internal standards shown. D–F Western blot analysis of SEC fractions from native tissue lysates of liver ( D ), kidney ( E ), and lung ( F ). Twelve-week-old male C57BL/6 mice were used. Lysates were fractionated on a Superose 6 Increase 3.2/300 column. Immunoblots probing PRPS complex members and internal standards are shown. Circular pictograms below SEC immunoblots schematize PRPS complex configurations; double circle denotes multiple copies of the protein interacting within the heteromeric complex. Coomassie staining ( A ) and western blot data ( C – F ) are representative of at least 3 biological repeats. Source data are provided as a Source Data file.

Techniques: SDS Page, Staining, Immunoprecipitation, Stable Transfection, Expressing, Control, Mass Spectrometry, Transformation Assay, Western Blot, Size-exclusion Chromatography

$A Western blot validating CRISPR-Cas9-generated isogenic knockout cell lines. B Proliferation of NIH3T3 parental and knockout cell lines generated in ( A ) ( n = 3 technical replicates). C Western blot validating HRAS G12V -overexpression in NIH3T3 parental and P2/AP1/AP2 KO cell lines. Phospho-MAPK (Erk1/2) (T202/Y204) used as a marker for activation of signaling pathways upon HRAS G12V overexpression. D Representative images from soft agar colony formation assay performed in NIH3T3 parental and P2/AP1/AP2 KO cells expressing HRAS G12V . Scale bar, 500 μm. E Quantification of colonies from ( D ) ( n = 3 experimental replicates). F Oxygen consumption rate (OCR) measured by Seahorse ATP Rate Assay in NIH3T3 parental and P2/AP1/AP2 KO cell lines ( n = 8 technical replicates). G OCR measured by Seahorse mitochondrial stress tests in NIH3T3 parental and NDI1-expressing P2/AP1/AP2 KO cell lines ( n = 7 technical replicates). H Proliferation of nucleoside supplemented and NDI1 expressing NIH3T3 P2/AP1/AP2 cell lines ( n = 3 technical replicates). I 13 C 6 -glucose metabolic labeling performed in NIH3T3 parental and P2/AP1/AP2 KO cell lines for 30 min and 5 h. Unlabeled, 13 C-labeled (30 min), and 13 C-labeled (5 h) samples are represented in brown, green, and blue colors, respectively. 13 C-enrichments quantified from 1 H-NMR spectra ( n = 3 experimental replicates). J Western blot analysis of SEC fractions collected from NIH3T3 P2/AP1/AP2 KO native whole-cell lysates. Cell lysates were fractionated on a Superose 6 Increase 3.2/300 column. In the pictogram below SEC immunoblots, the double circle with dotted inner circle denotes multiple copies of PRPS1 forming homo-oligomers. Data are represented as mean ± SD for ( B , E – H ) and mean ± SEM for ( I ). Statistical comparisons made using one way ANOVA followed by Tukey’s HSD post hoc test ( E , I ). Source data are provided as a Source Data file.$

Journal: Nature Communications

Article Title: The role of gene duplication and paralog specialisation in the evolution of the mammalian PRPS complex

doi: 10.1038/s41467-025-61216-z

Figure Lengend Snippet: A Western blot validating CRISPR-Cas9-generated isogenic knockout cell lines. B Proliferation of NIH3T3 parental and knockout cell lines generated in ( A ) ( n = 3 technical replicates). C Western blot validating HRAS G12V -overexpression in NIH3T3 parental and P2/AP1/AP2 KO cell lines. Phospho-MAPK (Erk1/2) (T202/Y204) used as a marker for activation of signaling pathways upon HRAS G12V overexpression. D Representative images from soft agar colony formation assay performed in NIH3T3 parental and P2/AP1/AP2 KO cells expressing HRAS G12V . Scale bar, 500 μm. E Quantification of colonies from ( D ) ( n = 3 experimental replicates). F Oxygen consumption rate (OCR) measured by Seahorse ATP Rate Assay in NIH3T3 parental and P2/AP1/AP2 KO cell lines ( n = 8 technical replicates). G OCR measured by Seahorse mitochondrial stress tests in NIH3T3 parental and NDI1-expressing P2/AP1/AP2 KO cell lines ( n = 7 technical replicates). H Proliferation of nucleoside supplemented and NDI1 expressing NIH3T3 P2/AP1/AP2 cell lines ( n = 3 technical replicates). I 13 C 6 -glucose metabolic labeling performed in NIH3T3 parental and P2/AP1/AP2 KO cell lines for 30 min and 5 h. Unlabeled, 13 C-labeled (30 min), and 13 C-labeled (5 h) samples are represented in brown, green, and blue colors, respectively. 13 C-enrichments quantified from 1 H-NMR spectra ( n = 3 experimental replicates). J Western blot analysis of SEC fractions collected from NIH3T3 P2/AP1/AP2 KO native whole-cell lysates. Cell lysates were fractionated on a Superose 6 Increase 3.2/300 column. In the pictogram below SEC immunoblots, the double circle with dotted inner circle denotes multiple copies of PRPS1 forming homo-oligomers. Data are represented as mean ± SD for ( B , E – H ) and mean ± SEM for ( I ). Statistical comparisons made using one way ANOVA followed by Tukey’s HSD post hoc test ( E , I ). Source data are provided as a Source Data file.

Techniques: Western Blot, CRISPR, Generated, Knock-Out, Over Expression, Marker, Activation Assay, Protein-Protein interactions, Soft Agar Assay, Expressing, Labeling

$A , B Western blot analysis of SEC fractions collected from native whole cell lysates of NIH3T3 P2/AP2 KO cells ( A ) and P2/AP1/AP2 KO cells stably expressing AP1 ( B ). C ALFA pulldown from whole cell extracts of NIH3T3 parental and P2/AP2 KO cells transiently transfected with PRPS1-ALFA. D–G Western blot analysis of SEC fractions collected from native whole-cell lysates of NIH3T3 AP1 KO cells ( D ), AP1/AP2 KO cells ( E ), AP1 KO cells ( F ), and AP1/AP2 KO cells ( G ). H Summary comparing SEC profiles from NIH3T3 parental and CRISPR KO lines. The x -axis represents relative molecular weight of the complex, and the y -axis represents the number of PRPS complex members interacting in the complex. Cell lysates for ( A , B , D , and E ) were fractionated on a Superose 6 Increase 3.2/300 column, while cell lysates for ( F ) and ( G ) were fractionated on a Yarra SEC-2000 column. Circular pictograms below SEC immunoblots schematize PRPS complex configurations. Double circle denotes multiple copies of the protein interacting in a heteromeric complex. Double circle with dotted inner circle denotes multiple copies forming homo-oligomers. Single circle denotes a single protein interacting within the complex. Circle with inner vertical lines denotes proteins forming a trimer or tetramer. Western blot data ( A – G ) are representative of at least 2 biological repeats. Source data are provided as a Source Data file.$

Journal: Nature Communications

Article Title: The role of gene duplication and paralog specialisation in the evolution of the mammalian PRPS complex

doi: 10.1038/s41467-025-61216-z

Figure Lengend Snippet: A , B Western blot analysis of SEC fractions collected from native whole cell lysates of NIH3T3 P2/AP2 KO cells ( A ) and P2/AP1/AP2 KO cells stably expressing AP1 ( B ). C ALFA pulldown from whole cell extracts of NIH3T3 parental and P2/AP2 KO cells transiently transfected with PRPS1-ALFA. D–G Western blot analysis of SEC fractions collected from native whole-cell lysates of NIH3T3 AP1 KO cells ( D ), AP1/AP2 KO cells ( E ), AP1 KO cells ( F ), and AP1/AP2 KO cells ( G ). H Summary comparing SEC profiles from NIH3T3 parental and CRISPR KO lines. The x -axis represents relative molecular weight of the complex, and the y -axis represents the number of PRPS complex members interacting in the complex. Cell lysates for ( A , B , D , and E ) were fractionated on a Superose 6 Increase 3.2/300 column, while cell lysates for ( F ) and ( G ) were fractionated on a Yarra SEC-2000 column. Circular pictograms below SEC immunoblots schematize PRPS complex configurations. Double circle denotes multiple copies of the protein interacting in a heteromeric complex. Double circle with dotted inner circle denotes multiple copies forming homo-oligomers. Single circle denotes a single protein interacting within the complex. Circle with inner vertical lines denotes proteins forming a trimer or tetramer. Western blot data ( A – G ) are representative of at least 2 biological repeats. Source data are provided as a Source Data file.

Techniques: Western Blot, Stable Transfection, Expressing, Transfection, CRISPR, Molecular Weight

$A–C Western blot analysis of SEC fractions collected from native whole-cell lysates of NIH3T3 AP1 KO cells stably expressing AP1 lacking the non-homologous region (NHR) ( A ), AP1/AP2 KO cells stably expressing chimeric AP1 with AP2’s N-terminus (residues 1–95) B and chimeric AP2 with AP1’s N-terminus (residues 1–95) ( C ). D Schematic representation of alternative start sites in mammalian AP1 and AP2 and their consequent translation into short and long isoforms. Base positions correspond to the mouse homologs. TSS1 and TSS2 represent upstream and downstream translation start sites, respectively. E Multiple isoforms for AP1 and AP2 detected under optimal SDS-PAGE resolution. Arrows in immunoblots indicate the longer isoforms of AP1 and AP2, corresponding to N-terminal leader sequences of 29 and 12 amino acids, respectively. F Phylogenetic distribution of PRPS homologs (PRPS1, PRPS2, PRPSAP2, PRPSAP2 with N-terminal leader sequence, PRPSAP1, and PRPSAP1 with N-terminal leader sequence) in chordates. Presence/absence noted across the tree. PRPSAP2 and PRPSAP1 isoforms with N-terminal leader sequences emerged in ancestors of Amniota and Osteichthyes, respectively. G–J Western blot analysis of SEC fractions collected from NIH3T3 AP1/AP2 KO cells stably expressing the short isoform of AP1 ( G ), long isoform of AP1 ( H ), short isoform of AP2 ( I ), and long isoform of AP2 ( J ). Cell lysates were fractionated on a Superose 6 Increase 3.2/300 column. Circular pictograms below SEC immunoblots schematize PRPS complex configurations. Double circle denotes multiple copies of the protein interacting in a heteromeric complex. Double circle with dotted inner circle denotes multiple copies forming homo-oligomers. Single circle denotes a single protein interacting within the complex. Circle with inner vertical lines denotes proteins forming a trimer or tetramer. Western blot data ( A – C , E , G – J ) are representative of at least 2 biological repeats. Source data are provided as a Source Data file.$

Journal: Nature Communications

Article Title: The role of gene duplication and paralog specialisation in the evolution of the mammalian PRPS complex

doi: 10.1038/s41467-025-61216-z

Figure Lengend Snippet: A–C Western blot analysis of SEC fractions collected from native whole-cell lysates of NIH3T3 AP1 KO cells stably expressing AP1 lacking the non-homologous region (NHR) ( A ), AP1/AP2 KO cells stably expressing chimeric AP1 with AP2’s N-terminus (residues 1–95) B and chimeric AP2 with AP1’s N-terminus (residues 1–95) ( C ). D Schematic representation of alternative start sites in mammalian AP1 and AP2 and their consequent translation into short and long isoforms. Base positions correspond to the mouse homologs. TSS1 and TSS2 represent upstream and downstream translation start sites, respectively. E Multiple isoforms for AP1 and AP2 detected under optimal SDS-PAGE resolution. Arrows in immunoblots indicate the longer isoforms of AP1 and AP2, corresponding to N-terminal leader sequences of 29 and 12 amino acids, respectively. F Phylogenetic distribution of PRPS homologs (PRPS1, PRPS2, PRPSAP2, PRPSAP2 with N-terminal leader sequence, PRPSAP1, and PRPSAP1 with N-terminal leader sequence) in chordates. Presence/absence noted across the tree. PRPSAP2 and PRPSAP1 isoforms with N-terminal leader sequences emerged in ancestors of Amniota and Osteichthyes, respectively. G–J Western blot analysis of SEC fractions collected from NIH3T3 AP1/AP2 KO cells stably expressing the short isoform of AP1 ( G ), long isoform of AP1 ( H ), short isoform of AP2 ( I ), and long isoform of AP2 ( J ). Cell lysates were fractionated on a Superose 6 Increase 3.2/300 column. Circular pictograms below SEC immunoblots schematize PRPS complex configurations. Double circle denotes multiple copies of the protein interacting in a heteromeric complex. Double circle with dotted inner circle denotes multiple copies forming homo-oligomers. Single circle denotes a single protein interacting within the complex. Circle with inner vertical lines denotes proteins forming a trimer or tetramer. Western blot data ( A – C , E , G – J ) are representative of at least 2 biological repeats. Source data are provided as a Source Data file.

Techniques: Western Blot, Stable Transfection, Expressing, SDS Page, Sequencing

A Chronological duplication events of a Class I PRPS encoding gene in indicated eukaryotes. Grey triangle traces the evolutionary trajectory leading to the emergence of mammalian PRPS homologs. Solid vertical lines within the triangle denote the origin of major lineages; dashed lines indicate significant duplication events giving rise to PRPS homologs, with notable functional innovations occurring post-duplication. Class II PRPS, likely present in the LECA, is lost in craniates, followed by rapid expansion of Class I PRPS homologs in jawed vertebrates. Left square bracket indicates a gene duplication event; left round bracket indicates an extension at the N-terminus. Molecular clock-based evolutionary timeline (in millions of years before present, Ma) adapted from ref. . B Association of different PRPS complex configurations with cellular fitness in mammals (i) PRPS1 alone undergoes self-assembly to form homo-oligomers. (ii) PRPS1 and AP1 cannot interact directly and form separate homo-oligomers. (iii) PRPS2 binds PRPS1 to form a dimer and disrupts PRPS1 homotypic assemblies. (iv) AP2 preferentially binds with PRPS1 to form a trimer/tetramer to nucleate the complex. (v) AP1 preferentially binds with AP2 and PRPS2 to elongate the PRPS complex via its N-terminus. (vi) The longer isoform of AP1 with flexible N-terminal extension likely caps the complex. The weight of the connectors between the circles representing proteins reflects the predicted strength of protein-protein interactions; heavier lines indicate stronger predicted interactions.

Journal: Nature Communications

Article Title: The role of gene duplication and paralog specialisation in the evolution of the mammalian PRPS complex

doi: 10.1038/s41467-025-61216-z

Figure Lengend Snippet: A Chronological duplication events of a Class I PRPS encoding gene in indicated eukaryotes. Grey triangle traces the evolutionary trajectory leading to the emergence of mammalian PRPS homologs. Solid vertical lines within the triangle denote the origin of major lineages; dashed lines indicate significant duplication events giving rise to PRPS homologs, with notable functional innovations occurring post-duplication. Class II PRPS, likely present in the LECA, is lost in craniates, followed by rapid expansion of Class I PRPS homologs in jawed vertebrates. Left square bracket indicates a gene duplication event; left round bracket indicates an extension at the N-terminus. Molecular clock-based evolutionary timeline (in millions of years before present, Ma) adapted from ref. . B Association of different PRPS complex configurations with cellular fitness in mammals (i) PRPS1 alone undergoes self-assembly to form homo-oligomers. (ii) PRPS1 and AP1 cannot interact directly and form separate homo-oligomers. (iii) PRPS2 binds PRPS1 to form a dimer and disrupts PRPS1 homotypic assemblies. (iv) AP2 preferentially binds with PRPS1 to form a trimer/tetramer to nucleate the complex. (v) AP1 preferentially binds with AP2 and PRPS2 to elongate the PRPS complex via its N-terminus. (vi) The longer isoform of AP1 with flexible N-terminal extension likely caps the complex. The weight of the connectors between the circles representing proteins reflects the predicted strength of protein-protein interactions; heavier lines indicate stronger predicted interactions.

Techniques: Functional Assay, Protein-Protein interactions

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