Evidence map›Paper›PMID 35181508›Full record

ArticleProtein expression and purification2022

Expression and production of pigment epithelium-derived factor (PEDF) and PEDF receptor variants from mammalian and bacterial cells.

Jeanee Bullock, Federica Polato, Daniel C Lulli, Vatsala Sagar, Laura Abaandou, Joseph Shiloach, S Patricia Becerra

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Protein expression and purification, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.

0numbers the graph read from it
0cells of the map it votes in
3citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

3 citing papers in PubMed.

  1. Review
  2. Article
  3. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

7 authors.

Jeanee BullockSection of Protein Structure and Function-LRCMB, National Eye Institute, National Institutes of Health, Bethesda, MD, USA; Department of Biochemistry and Molecular & Cellular Biology, Georgetown University Medical Center, Washington, DC, USA.
Federica PolatoSection of Protein Structure and Function-LRCMB, National Eye Institute, National Institutes of Health, Bethesda, MD, USA.
Daniel C LulliSection of Protein Structure and Function-LRCMB, National Eye Institute, National Institutes of Health, Bethesda, MD, USA.
Vatsala SagarSection on Molecular Structure and Functional Genomics, National Eye Institute, National Institutes of Health, Bethesda, MD, USA.
Laura AbaandouBiotechnology Core Laboratory, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA; Department of Chemistry and Biochemistry, George Mason University, Fairfax, VA, USA.
Joseph ShiloachBiotechnology Core Laboratory, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA.
S Patricia BecerraSection of Protein Structure and Function-LRCMB, National Eye Institute, National Institutes of Health, Bethesda, MD, USA. Electronic address: becerrap@nei.nih.gov.

Funding

Role of Protein Interactions in Retina Development and FunctionZIAEY000306 · NEI · NATIONAL EYE INSTITUTE · PI BECERRA, S. PATRICIA · 2009 to 2025
$15.6M
Production and purification of biological compoundsZICDK015500 · NIDDK · NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES · PI SHILOACH, JOSEPH · 2009 to 2022
$7.2M
ROLE OF PROTEIN INTERACTIONS IN RETINA DEVELOPMENT AND FUNCTIONZ01EY000306 · NEI · NATIONAL EYE INSTITUTE · PI BECERRA, SOFIA P · 1995 to 2008
$2.3M
Large-scale production and purification of biological compoundsZIBDK015500 · NIDDK · NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES · PI SHILOACH, JOSEPH · 2015 to 2016
$1.2M
LARGE-SCALE PRODUCTION &PURIFICATION OF COMPOUNDS WITH BIOLOGICAL ACTIVITYZ01DK015500 · NIDDK · NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES · PI SHILOACH, JOSEPH · 1986 to 2008
$568k
Intramural NIH HHS Z01 DK015500Intramural NIH HHS Z01 EY000306Intramural NIH HHS ZIA EY000306Intramural NIH HHS ZIB DK015500Intramural NIH HHS ZIC DK015500
6 · The paper itself

Abstract

Human SERPINF1 gene codes for pigment epithelium-derived factor (PEDF), a secreted glycoprotein and member of the SERPIN superfamily. To obtain large amounts of recombinant PEDF proteins, we subcloned the coding sequence of human SERPINF1 mutated versions into the pCEP4 vector and generated stably transfected HEK.Ebna cells. The cells produced and secreted recombinant PEDF proteins into the culturing media. The recombinant PEDF proteins were purified by ion-exchange column chromatography and milligram amounts of highly purified protein were recovered. PEDF has affinity for PEDF-receptor (PEDF-R), a membrane-linked lipase encoded by the PNPLA2 gene. Recombinant PEDF-R truncated versions were obtained from Escherichia coli containing expression vectors with human PNPLA2 cDNAs with 3'end deletions and by induction with isopropyl β-d-1-thiogalactopyranoside. The bacterially derived PEDF-R proteins in insoluble inclusion bodies were solubilized with urea and purified by cation-exchange column chromatography. C-terminally truncated PEDF-R versions containing the ligand binding region retained the ability to bind PEDF. The data demonstrate that mammalian-derived recombinant PEDF and bacterially derived recombinant PEDF-R can be produced and purified in large amounts for further use in structural and biological studies.

Indexed as

SerpinsAnimalsEye ProteinsHumansMammalsNerve Growth FactorsPigment Epithelium-Derived FactorReceptors, NeuropeptideRecombinant ProteinsEye ProteinsNerve Growth FactorsPigment Epithelium-Derived Factorpigment epithelium-derived factor receptorReceptors, NeuropeptideRecombinant ProteinsSerpinsBacterial overexpressionMammalian overexpressionPEDFPNPLA2PurificationSerpin

Identifiers

PMID35181508
PMCPMC8934279

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.