Structural modelling of human complement FHR1 and two of its synthetic derivatives provides insight into their in-vivo functions

Ruiz-Molina, Natalia; Parsons, Juliana; Decker, Eva L.; Reski, Ralf

doi:10.1016/j.csbj.2023.02.002

Cited by 3 publications

(3 citation statements)

References 86 publications

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“…While most production hosts are vascular plants, such as Nicotiana benthamiana (Nicotiana) and Daucus carota (carrot), the non-vascular moss Physcomitrella has a proven track-record for the production of pharmaceuticals and bioactive ingredients (Decker and Reski, 2020; Verdú-Navarro et al, 2023; Munoz et al, 2024). In our attempts to constantly optimize this moss for molecular farming approaches (Reski et al, 2015; Ruiz-Molina et al, 2023), we concentrate on gene expression (e.g., Top et al, 2021; Niederau et al, 2024), bioproduction (e.g., Ruiz-Molina et al, 2022), and glycoengineering (e.g., Decker and Reski, 2012; Bohlender et al, 2022). In the latter field, plant-typical glyco-structures have been abolished (Parsons et al, 2012), and stable in-vivo protein sialylation has been achieved (Bohlender et al, 2020).…”

Section: Discussionmentioning

confidence: 99%

Differential prolyl hydroxylation by six Physcomitrella prolyl-4 hydroxylases

Rempfer,

Hoernstein,

van Gessel

et al. 2024

Preprint

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The hydroxylation of proline residues to 4-trans-hydroxyproline (Hyp) is a common post-translational protein modification in plants, mediated by prolyl 4-hydroxylases (P4Hs). Hyps predominantly occur in a group of cell wall proteins, the Hydroxyproline-rich glycoproteins (HRGPs), where they are frequently O-glycosylated. While prolyl-hydroxylation and O-glycosylation are important, e.g. for cell wall stability, they are undesired on plant-made pharmaceuticals. Sequence motifs recognized for prolyl-hydroxylation derived from vascular plants were proposed but did not include data from mosses, such as Physcomitrella. Here, a phylogenetic reconstruction of plant P4Hs identified six P4Hs in four subfamilies in mosses. We analysed the amino acid sequences and structural environments around Hyps in Physcomitrella utilizing 73 Hyp sites in 24 secretory proteins from multiple MS/MS datasets, and found that prolines in close proximity to other prolines, alanine, serine, threonine and valine were preferentially hydroxylated. About 95% of the Hyp sites were predictable with a combination of previously defined motifs and methods. In our data, AOV (Ala-Hyp-Val) was the most frequent prolyl-hydroxylation pattern. Additionally, short arabinose chains were attached to Hyps in two cell-wall pectinesterases. A combination of 443 AlphaFold structure models and our MS data of peptides with nearly 3000 proline sites found Hyps predominantly on protein surfaces in disordered regions. Moss-produced human erythropoietin (EPO) exhibited plant-specific O-glycosylation with arabinose chains on two Hyps. This modification was significantly reduced in a P4H1 single knock-out (KO) Physcomitrella mutant. Quantitative proteomics after isotope labelling with different P4H-KO moss mutants revealed specific changes in the amount of proteins, including HRGPs, and modified prolyl-hydroxylation pattern from the mutants, suggesting a differential function of the six Physcomitrella P4Hs. Quantitative RT-PCR proved a differential effect of single P4H KOs on the expression of the other five p4h genes, suggesting a partial compensation of the mutation. AlphaFold-Multimer models for Physcomitrella P4H1 and its target EPO peptide superposed with the crystal structure of Chlamydomonas P4H1 and a peptide substrate suggested significant amino acids in the active centre of the enzyme that form H-bonds with the peptide substrate, and revealed differences between P4H1 and the other five Physcomitrella P4Hs.

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Section: Discussionmentioning

confidence: 99%

Differential prolyl hydroxylation by six Physcomitrella prolyl-4 hydroxylases

Rempfer,

Hoernstein,

van Gessel

et al. 2024

Preprint

Self Cite

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“…While most production hosts are vascular plants, such as Nicotiana benthamiana (Nicotiana) and Daucus carota (carrot), the non-vascular moss Physcomitrella has a proven track-record for the production of pharmaceuticals and bioactive ingredients [119] , [20] , [75] . In our attempts to constantly optimize this moss for molecular pharming [90] , [92] , we concentrate on gene expression [107] , [78] , bioproduction [93] , and glycoengineering [19] , [6] . In the latter field, plant-typical glyco-structures have been abolished [82] , and stable in-vivo protein sialylation has been achieved [5] .…”

Section: Discussionmentioning

confidence: 99%

Differential prolyl hydroxylation by six Physcomitrella prolyl-4 hydroxylases

Rempfer,

Hoernstein,

van Gessel

et al. 2024

Computational and Structural Biotechnology Journal

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“…Figure 4 shows schematic changes in N-and O-glycosylation accompanying tumour formation and progression. Sialylation and hyper sialylation often lead to immune system processes' modulation, as in the case of factor H recognising sialoconjugates and modulating complement activation [67], or the presenc of α2,8-Sia containing structures protecting a tumour from the cytotoxic effects of NK (natural killer) cells [68]. Truncated sialylated O-glycans, on the other hand, were pro posed as a target for novel anti-cancer immunotherapy; however, the main obstacle in thi area is a low specificity and cross-reactivity [69].…”

Section: Three Levels Of Regulation-genes Substrates and Golgimentioning

confidence: 99%

Medical Relevance, State-of-the-Art and Perspectives of “Sweet Metacode” in Liquid Biopsy Approaches

Pinkeova,

Kosutova,

Jane

et al. 2024

Diagnostics

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This review briefly introduces readers to an area where glycomics meets modern oncodiagnostics with a focus on the analysis of sialic acid (Neu5Ac)-terminated structures. We present the biochemical perspective of aberrant sialylation during tumourigenesis and its significance, as well as an analytical perspective on the detection of these structures using different approaches for diagnostic and therapeutic purposes. We also provide a comparison to other established liquid biopsy approaches, and we mathematically define an early-stage cancer based on the overall prognosis and effect of these approaches on the patient’s quality of life. Finally, some barriers including regulations and quality of clinical validations data are discussed, and a perspective and major challenges in this area are summarised.

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Structural modelling of human complement FHR1 and two of its synthetic derivatives provides insight into their in-vivo functions

Cited by 3 publications

References 86 publications

Differential prolyl hydroxylation by six Physcomitrella prolyl-4 hydroxylases

Differential prolyl hydroxylation by six Physcomitrella prolyl-4 hydroxylases

Differential prolyl hydroxylation by six Physcomitrella prolyl-4 hydroxylases

Medical Relevance, State-of-the-Art and Perspectives of “Sweet Metacode” in Liquid Biopsy Approaches

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