Recombinant Human Proinsulin Protein, CF

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human Proinsulin Protein, CF Summary

• Details of Functionality: Measured in a serum-free cell proliferation assay using MCF‑7 human breast cancer cells. Karey, K.P. et al. (1988) Cancer Research 48:4083. The ED₅₀ for this effect is 0.15-0.75 μg/mL.
• Source: E. coli-derived human Proinsulin protein
  Phe25-Asn110, with an N-terminal Met, 6-His tag and Lys
• Accession #: NP_000198
• N-terminal Sequence: Met
• Protein/Peptide Type: Recombinant Proteins
• Gene: INS
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <0.01 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 10.5 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 9 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Reconstitution Instructions: Reconstitute at 100 μg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Proinsulin Protein, CF

• IDDM2
• ILPR
• insulin
• IRDN
• MODY10
• Proinsulin

Background

Proinsulin is synthesized as a single chain, 110 amino acid (aa) preproprecursor that contains a 24 aa signal sequence and an 86 aa proinsulin propeptide. Following removal of the signal peptide, the proinsulin peptide undergoes further proteolysis to generate mature insulin, a 51 aa disulfide-linked dimer that consists of a 30 aa B chain (aa 25 - 54) bound to a 21 aa A chain (aa 90 - 110). The 34 aa intervening peptide (aa 55 - 89) that connects the B and A chains is termed the C-peptide. Human proinsulin shares 84% and 80% aa sequence identity with rat and bovine proinsulin, respectively. Most of the sequence variation between species occurs in the region of the C-peptide (1). This peptide generates a structural conformation that allows for the correct formation of the intrachain disulphide bonds (1). Insulin is a molecule that facilitates the cellular uptake of glucose. This is accomplished by regulating the appearance of membrane glucose transporters. Low insulin levels or lack of insulin are associated with type 2 and type 1 diabetes mellitus, respectively. These conditions are associated with an increased risk for microvascular complications such as retinopathy, nephropathy, and peripheral neuropathy (3). Proinsulin also circulates, but its physiologic role is less well understood. It does possess about 25% of the activity of mature insulin, but it would seem unlikely to be a natural substitute for insulin (4). In type 2 diabetes, an elevated proinsulin to insulin ratio in the circulation is a well-known abnormality (5 - 9). Perhaps this abnormality represents either compromised proteolytic processing or a general inability to process increased levels of insulin precursor (5). In any event, proinsulin will stimulate amylin secretion by beta -cells, and amyloid formation in pancreatic islets that promotes decreased beta cell function (10). Studies also suggest that fasting serum proinsulin may be a better predictor of future type 2 diabetes than fasting insulin levels in obese children (11).

1. Bell, G.I. et al. (1980) Nature 284:26.
2. Barbetti, F. et al. (1990) J. Clin. Endocrinol. Metab. 71:164.
3. Forst, T. et al. (2008) Exp. Diabetes Res. 2008:176245.
4. Steffes, M.W. et al. (2003) Diabetes Care 26:832.
5. Roder, M.E. et al. (1999) Diabetes Care 22:609.
6. Porte, D. Jr. (1991) Diabetes 40:166.
7. Gordon, P. et al. (1974) Diabetologia 34:483.
8. Saad, M.F. et al. (1990) J. Clin. Endocrinol. Metab. 70:1247.
9. Roder, M.E. et al. (1995) J. Clin. Endocrinol. Metab. 80:2359.
10. Dworacka, M. et al. (2006) Int. J. Clin. Pharmacol. Ther. 44:14.
11. Kamoda, T. et al. (2006) Diabetes Obes. Metab. 8:192.

Publications for Proinsulin (1336-PN)(10)

Publications using 1336-PN

• DT Meier, L Rachid, SJ Wiedemann, S Traub, K Trimiglioz, M Stawiski, L Sauteur, DV Winter, C Le Foll, C Brégère, R Guzman, A Odermatt, M Böni-Schne, MY Donath Prohormone convertase 1/3 deficiency causes obesity due to impaired proinsulin processing Nature Communications, 2022-08-13;13(1):4761. 2022-08-13 [PMID: 35963866] (In Vivo, Mouse)
• J Viviano, M Brecker, C Ferrara-Co, L Suaud, RC Rubenstein ERp29 as a regulator of Insulin biosynthesis PLoS ONE, 2020-05-20;15(5):e0233502. 2020-05-20 [PMID: 32433667] (Western Blot Control, Mouse)
• PL Toledo, JM Torkko, A Müller, C Wegbrod, A Sönmez, M Solimena, MR Ermácora ICA512 RESP18 homology domain is a protein condensing factor and insulin fibrillation inhibitor J. Biol. Chem., 2019-04-12;0(0):. 2019-04-12 [PMID: 30979722] (Bioassay, Human)
• Y Liu, HY Wang, L Zhou, Y Su, WC Shen Biodistribution, activation, and retention of proinsulin-transferrin fusion protein in the liver: Mechanism of liver-targeting as an insulin prodrug J Control Release, 2018-02-20;275(0):186-191. 2018-02-20 [PMID: 29471040] (Cell Culture, Mouse)
• YJ Hsieh, KY Chien, IF Yang, IN Lee, CC Wu, TY Huang, JS Yu Oxidation of protein-bound methionine in Photofrin-photodynamic therapy-treated human tumor cells explored by methionine-containing peptide enrichment and quantitative proteomics approach Sci Rep, 2017-05-02;7(1):1370. 2017-05-02 [PMID: 28465586] (Bioassay)
• Farino Z, Morgenstern T, Vallaghe J, Gregor N, Donthamsetti P, Harris P, Pierre N, Freyberg R, Charrier-Savournin F, Javitch J, Freyberg Z Development of a Rapid Insulin Assay by Homogenous Time-Resolved Fluorescence. PLoS ONE, 2016-02-05;11(2):e0148684. 2016-02-05 [PMID: 26849707] (ELISA (Standard), Rat)
• Rulifson I, Cao P, Miao L, Kopecky D, Huang L, White R, Samayoa K, Gardner J, Wu X, Chen K, Tsuruda T, Homann O, Baribault H, Yamane H, Carlson T, Wiltzius J, Li Y Identification of Human Islet Amyloid Polypeptide as a BACE2 Substrate. PLoS ONE, 2016-02-03;11(2):e0147254. 2016-02-03 [PMID: 26840340] (Bioassay, Human)
• Mbongue J, Nicholas D, Zhang K, Kim N, Hamilton B, Larios M, Zhang G, Umezawa K, Firek A, Langridge W Induction of indoleamine 2, 3-dioxygenase in human dendritic cells by a cholera toxin B subunit-proinsulin vaccine. PLoS ONE, 2015-02-25;10(2):e0118562. 2015-02-25 [PMID: 25714914] (Bioassay, Human)
• Nardi F, Lipina C, Magill D, Hage Hassan R, Hajduch E, Gray A, Hundal H Enhanced insulin sensitivity associated with provision of mono and polyunsaturated fatty acids in skeletal muscle cells involves counter modulation of PP2A. PLoS ONE, 2014-03-14;9(3):e92255. 2014-03-14 [PMID: 24632852] (Bioassay, Human)
• Wang , Yan, Shao , Juntang, Zaro , Jennica, Shen , Wei-Chia Proinsulin-transferrin fusion protein as a novel long-acting insulin analog for the inhibition of hepatic glucose production. Diabetes, 2013-12-18;63(5):1779-88. 2013-12-18 [PMID: 24353179] (In Vivo, Mouse)

Bioinformatics

• Gene Symbol: INS
• Uniprot:
  • NP_000198(Human)