Human Insulin from Recombinant DNA Technology

Johnson, Irving S.

doi:10.1126/science.6337396

Cited by 317 publications

(142 citation statements)

References 7 publications

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“…Recombinant human insulin was obtained from the laboratories of Merck & Co. RHI is a commercially available material; a generic description for the bioprocess used in preparing it can be found in the paper by Johnson 35 . A stock solution (5.6 mg/ml, 0.969 mM) was made in HBS (10 mM Hepes pH=7.4, 150 mM NaCl) and stored at 4°C.…”

Section: Methodsmentioning

confidence: 99%

Structure of the insulin receptor–insulin complex by single-particle cryo-EM analysis

Scapin

Dandey

Zhang

et al. 2018

Nature

192

243

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Summary The insulin receptor (IR) is a dimeric protein that plays a crucial role in controlling glucose homeostasis, regulating lipid, protein and carbohydrate metabolism, and modulating brain neurotransmitter levels1,2. IR dysfunctions have been associated with many diseases, including diabetes, cancer, and Alzheimer’s1,3,4. The primary sequence has been known since the 1980s5, and is composed of an extracellular portion (ectodomain, ECD), a single transmembrane helix and an intracellular tyrosine kinase domain. Insulin binding to the dimeric ECD triggers kinase domain auto-phosphorylation and subsequent activation of downstream signaling molecules. Biochemical and mutagenesis data have identified two putative insulin binding sites (S1 and S2)6. While insulin bound to an ECD fragment containing S1 and the apo ectodomain have been characterized structurally7,8, details of insulin binding to the full receptor and the signal propagation mechanism are still not understood. Here we report single particle cryoEM reconstructions for the 1:2 (4.3 Å) and 1:1 (7.4 Å) IR ECD dimer:Insulin complexes. The symmetric 4.3 Å structure shows two insulin molecules per dimer, each bound between the Leucine-rich sub domain L1 of one monomer and the first fibronectin-like domain (FnIII-1) of the other monomer, and making extensive interactions with the α subunit C-terminal helix (α-CT helix). The 7.4 Å structure has only one similarly bound insulin per receptor dimer. The structures confirm the S1 binding interactions and define the full S2 binding site. These insulin receptor states suggest that recruitment of the α-CT helix upon binding of the first insulin changes the relative sub domain orientations and triggers downstream signal propagation.

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Section: Methodsmentioning

confidence: 99%

Structure of the insulin receptor–insulin complex by single-particle cryo-EM analysis

Scapin

Dandey

Zhang

et al. 2018

Nature

192

243

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“…Human insulin is prepared with this method by inserting the insulin gene into the E. Coli bacterial cell. This way, the implanted DNA induces the cell to produce insulin [62,63].…”

Section: Recombinant Dna Synthesismentioning

confidence: 99%

Purity profiling of Peptide Drugs

Verbeken¹

2012

JBABM

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The quality of a peptide drug mainly depends on its impurity profile, with the emphasis on the related impurities. These impurities may be biomedically active, alter the desired efficacy or induce unwanted toxicity, an aspect which is termed the "functional quality" of the peptide drug. Therefore, regulatory authorities have set up guidances or have legally established specification limits to assure a consistent purity of these peptide drugs. For the active pharmaceutical ingredients (APIs), the pharmacopoeial monographs are legally binding. Additional information can be found in regional and international guidelines. For the finished pharmaceutical drug products (FDPs) containing peptide active ingredients, only general guidelines are available. The construction of a complete related-impurity profile is very challenging due to the wide availability of different protecting groups, coupling agents and additives that may be used during peptide synthesis. In addition, chemical degradation, occurring during synthesis, formulation or at storage, may occur as well, including not only so-called pure chemical degradation but interaction with excipients as well. This review provides an update of the regulatory and scientific rationales behind the related impurities in peptide drugs.

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“…Roughly speaking, the challenge was to engineer a genetic cassette to overexpress a single gene product that was subsequently purified from the E. coli cell. The result was an insulin supply free of dangerous porcine-related antigens and a vivid demonstration of the potential for using genetic engineering to address health needs (Johnson 1983).…”

mentioning

confidence: 99%

Design-driven, multi-use research agendas to enable applied synthetic biology for global health

Carothers

2013

Syst Synth Biol

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Many of the synthetic biological devices, pathways and systems that can be engineered are multi-use, in the sense that they could be used both for commerciallyimportant applications and to help meet global health needs. The on-going development of models and simulation tools for assembling component parts into functionally-complex devices and systems will enable successful engineering with much less trial-and-error experimentation and laboratory infrastructure. As illustrations, I draw upon recent examples from my own work and the broader Keasling research group at the University of California Berkeley and the Joint BioEnergy Institute, of which I was formerly a part. By combining multi-use synthetic biology research agendas with advanced computer-aided design tool creation, it may be possible to more rapidly engineer safe and effective synthetic biology technologies that help address a wide range of global health problems.

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Human Insulin from Recombinant DNA Technology

Cited by 317 publications

References 7 publications

Structure of the insulin receptor–insulin complex by single-particle cryo-EM analysis

Structure of the insulin receptor–insulin complex by single-particle cryo-EM analysis

Purity profiling of Peptide Drugs

Design-driven, multi-use research agendas to enable applied synthetic biology for global health

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