Skip to main content
  • More from ADA
    • Diabetes Care
    • Clinical Diabetes
    • Diabetes Spectrum
    • ADA Standards of Medical Care in Diabetes
    • ADA Scientific Sessions Abstracts
    • BMJ Open Diabetes Research & Care
  • Subscribe
  • Log in
  • My Cart
  • Follow ada on Twitter
  • RSS
  • Visit ada on Facebook
Diabetes

Advanced Search

Main menu

  • Home
  • Current
    • Current Issue
    • Online Ahead of Print
    • ADA Scientific Sessions Abstracts
  • Browse
    • By Topic
    • Issue Archive
    • Saved Searches
    • ADA Scientific Sessions Abstracts
    • Diabetes COVID-19 Article Collection
    • Diabetes Symposium 2020
  • Info
    • About the Journal
    • About the Editors
    • ADA Journal Policies
    • Instructions for Authors
    • Guidance for Reviewers
  • Reprints/Reuse
  • Advertising
  • Subscriptions
    • Individual Subscriptions
    • Institutional Subscriptions and Site Licenses
    • Access Institutional Usage Reports
    • Purchase Single Issues
  • Alerts
    • E­mail Alerts
    • RSS Feeds
  • Podcasts
    • Diabetes Core Update
    • Special Podcast Series: Therapeutic Inertia
    • Special Podcast Series: Influenza Podcasts
    • Special Podcast Series: SGLT2 Inhibitors
    • Special Podcast Series: COVID-19
  • Submit
    • Submit a Manuscript
    • Submit Cover Art
    • ADA Journal Policies
    • Instructions for Authors
    • ADA Peer Review
  • More from ADA
    • Diabetes Care
    • Clinical Diabetes
    • Diabetes Spectrum
    • ADA Standards of Medical Care in Diabetes
    • ADA Scientific Sessions Abstracts
    • BMJ Open Diabetes Research & Care

User menu

  • Subscribe
  • Log in
  • My Cart

Search

  • Advanced search
Diabetes
  • Home
  • Current
    • Current Issue
    • Online Ahead of Print
    • ADA Scientific Sessions Abstracts
  • Browse
    • By Topic
    • Issue Archive
    • Saved Searches
    • ADA Scientific Sessions Abstracts
    • Diabetes COVID-19 Article Collection
    • Diabetes Symposium 2020
  • Info
    • About the Journal
    • About the Editors
    • ADA Journal Policies
    • Instructions for Authors
    • Guidance for Reviewers
  • Reprints/Reuse
  • Advertising
  • Subscriptions
    • Individual Subscriptions
    • Institutional Subscriptions and Site Licenses
    • Access Institutional Usage Reports
    • Purchase Single Issues
  • Alerts
    • E­mail Alerts
    • RSS Feeds
  • Podcasts
    • Diabetes Core Update
    • Special Podcast Series: Therapeutic Inertia
    • Special Podcast Series: Influenza Podcasts
    • Special Podcast Series: SGLT2 Inhibitors
    • Special Podcast Series: COVID-19
  • Submit
    • Submit a Manuscript
    • Submit Cover Art
    • ADA Journal Policies
    • Instructions for Authors
    • ADA Peer Review
Brief Genetics Report

Diabetes-Associated Mutations in Insulin Identify Invariant Receptor Contacts

  1. Bin Xu1,
  2. Shi-Quan Hu2,
  3. Ying-Chi Chu2,
  4. Shuhua Wang2,
  5. Run-ying Wang2,
  6. Satoe H. Nakagawa3,
  7. Panayotis G. Katsoyannis2 and
  8. Michael A. Weiss1
  1. 1Department of Biochemistry, Case Western Reserve School of Medicine, Cleveland, Ohio
  2. 2Department of Pharmacology and Biological Chemistry, Mount Sinai School of Medicine, New York University, New York, New York
  3. 3Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, Illinois
  1. Address correspondence and reprint requests to Michael A. Weiss, Case Western Reserve University, Department of Biochemistry, 10900 Euclid Ave., SOM Room W427, Cleveland, OH 44106-4935. E-mail: michael.weiss{at}case.edu
Diabetes 2004 Jun; 53(6): 1599-1602. https://doi.org/10.2337/diabetes.53.6.1599
PreviousNext
  • Article
  • Figures & Tables
  • Info & Metrics
  • PDF
Loading

Abstract

Mutations in human insulin cause an autosomal-dominant syndrome of diabetes and fasting hyperinsulinemia. We demonstrate by residue-specific photo cross-linking that diabetes-associated mutations occur at receptor-binding sites. The studies use para-azido-phenylalanine, introduced at five sites by total protein synthesis. Because two such sites (ValA3 and PheB24) are largely buried in crystal structures of the free hormone, their participation in receptor binding is likely to require a conformational change to expose a hidden functional surface. Our results demonstrate that this surface spans both chains of the insulin molecule and includes sites of rare human mutations that cause diabetes.

  • DTT, dithiothreitol
  • FnIII1, second fibronectin-homology domain
  • ID, insert domain
  • Pap, para-azido-phenylalanine
  • Phe, phenylalanine

The insulinopathies describe a monogenic form of adult-onset diabetes due to mutations in the insulin gene (1,2). Patients respond normally to exogenous insulin but exhibit fasting mutant hyperinsulinemia due to delayed receptor-mediated clearance of the variant hormone (2). Inheritance is autosomal dominant with variable penetrance. The presence of one wild-type allele permits normal growth and development; homozygous or hemizygous mutations have not been observed and are presumably incompatible with life. Diabetes-associated mutations may either impair binding of the variant insulin to its receptor or perturb subcellular trafficking and processing of the variant proinsulin in the pancreatic β-cell (2). Mutations that impair binding have been identified at three invariant sites: ValA3 → Leu, PheB24 → Ser, and PheB25 → Leu. By analogy to the nomenclature describing abnormal hemoglobins, these are designated insulins Wakayama, Los Angeles, and Chicago, respectively (1). We demonstrate here that these mutations occur at contact sites between insulin and the α-subunit of the insulin receptor.

The structure of insulin is well characterized by crystallography (3) and nuclear magnetic resonance spectroscopy (4,5) (Fig. 1A). Residues A3, B24, and B25 exhibit distinct environments. Whereas PheB25 projects from the surface, ValA3 and PheB24 are engaged in long-range interactions (Fig. 2). ValA3 contacts TyrB26 and ProB28 at an interface between the NH2-terminal A-chain α-helix and COOH-terminal B-chain β-strand (Fig. 2B). PheB24 packs against ValB12, LeuB15, TyrB16, and CysB19 to stabilize the supersecondary structure of the B-chain. In dimers and hexamers, PheB24 and PheB25 also participate in an intermolecular β-sheet, an essential element of insulin’s storage form in the β-cell (3). Whereas considerable evidence indicates that the exposed side chain of PheB25 contacts the insulin receptor (6, including previous photo cross-linking studies [7,8]), the roles of ValA3 and PheB24 have long been the subject of speculation (3,9–13).

To test whether residues A3, B24, and B25 contact the insulin receptor, we have synthesized insulin analogs containing a photo-activatable derivative of phenylalanine (Phe), para-azido-Phe (Pap) (8,14). Pap was chosen based on its rigidity and small size (relative to other photoactivable moieties), thus limiting the distance range for cross-linking. Modified A- and B-chains were prepared by solid-phase synthesis using the photostable precursor para-amino-Phe. To enable efficient detection of cross-linked peptides, the α-amino group of the B-chain was biotinylated (8). The nonstandard side chain was introduced into an engineered insulin monomer (DKP-insulin, which contains three B-chain substitutions: HisB10→Asp, ProB28→Lys, and LysB29→Pro), chosen as a template for its efficiency of synthesis, enhanced receptor binding, and absence of confounding self-association (4). A3, B24, and B25 para-amino-Phe analogs exhibit respective receptor-binding affinities of 2.0 ± 0.2, 59 ± 2, and 147 ± 3% relative to native insulin (Kd 0.48 nmol/l); the affinity of the biotin adduct of DKP-insulin is 132 ± 5% (assays performed in triplicate). Corresponding analogs were prepared at terminal positions A1 and A21 (ordinarily conserved as Gly and Asn, respectively) at the periphery of insulin’s putative receptor-binding surface (3,15); their relative receptor-binding affinities are 46 ± 5 and 79 ± 22%. AsnA21 projects from the surface near GlyB23 and PheB25 (Fig. 2A). Conversion of para-amino-Phe to Pap in the intact hormones was verified by mass spectrometry.

A3, B24, and B25 Pap analogs each exhibit rapid and efficient cross-linking to the ectodomain of the receptor on ultraviolet irradiation (red asterisks in Figs. 1B and C). Efficiency (defined as the probability of photo cross-linking of the Pap derivative once bound to the receptor) is highest for PapA3. Similar results are obtained with the lectin-purified holoreceptor. No covalent complex is observed in the absence of irradiation or in control studies of the para-amino-PheB25 precursor. Photo cross-linking is successively diminished by the addition of native insulin or by higher concentrations of IGF-1 (Fig. 1D). In contrast to the cross-linking at sites of clinical mutation, photo cross-linking of Pap derivatives at A1 and A21 is markedly less efficient (green arrows in lanes 6 and 14 in Fig. 1C; relative to A3, complexes are reduced by 19- and 4-fold, respectively). As a first step in identifying sites of cross-linking in the insulin receptor, we used partial proteolysis with trypsin and chymotrypsin to characterize fragments of the receptor α-subunit covalently bound to insulin. Analysis of such fragments demonstrates that PapB24 contacts the NH2-terminal L1 β-helix domain, the major hormone-binding region of the receptor (13). By contrast, PapB25 contacts the COOH-terminal region of the α-subunit in accordance with the pioneering study of Kurose et al. (8). Edman sequencing demonstrated that PapB25 cross-links to tryptic peptide 704-718 in the insert-domain tail. (The COOH-terminal residue of the α-subunit is 731.) In our hands, limited chymotryptic digestion of the covalent hormone-receptor complex, followed by reduction with dithiothreitol (DTT), yields a 34-kDa adduct that, on further digestion, yields a 20-kDa adduct. Following enzymatic deglycosylation, the apparent mass of this fragment is 14 kDa and thus contains about 120 amino acids. The results of Kurose et al. imply that the latter fragment contains the COOH-terminal portion of the α-subunit derived from the second fibronectin-homology domain (FnIII1) and insert domain (ID) (13).

PapA3, which is not predicted to contact the receptor in a current model based on electron-microscopic image reconstruction (16), cross-links the COOH-terminal to the L1 and cysteine-rich domains. To localize this site more precisely, a second PapA3 derivative was prepared in which the biotin tag was attached at the NH2-terminus of the A-chain (rather than the NH2-terminus of the B-chain; see research design and methods). This design facilitates mapping following DTT reduction as above. Limited chymotryptic digestion thus demonstrates that PapA3 cross-links to the same COOH-terminal 34-kDa and 20-kDa adducts as PapB25, i.e., within the FnIII1-ID–derived tail. We suggest that the A3 binding site (like that of PapB25 [8]) resides within the ID-derived portion, since the FnIII1 moiety may be deleted in active fragments of the α-subunit (13). Because the L1 domain and the COOH-terminal domain are distant in the sequence of the α-subunit, the present results suggest that these and other sites of photo cross-linking are nearby in the three-dimensional structure of the hormone-receptor complex (13,16). It is not known whether PapA3 and PapB25 cross-link to the same α-subunit or to dimer-related α-subunits within the α2β2 heterotetramer. Sites of weak cross-linking by PapA1 and PapA21 derivatives were not characterized.

Photo cross-linking of PapA3 and PapB24 derivatives is of structural interest. Because ValA3 and PheB24 are largely buried in crystal structures of insulin (3), it has been unclear whether these residues contact the receptor or serve as structural supports. A possible role for PheB24 in redirecting the main chain of insulin on receptor binding has been proposed based on the unexpectedly high activities of d-amino acid substitutions (9). We and others have hypothesized that detachment or reorganization of the COOH-terminal region of the B-chain near B24 exposes the side chains of PheB24 and ValA3 and thus enables them to contact the receptor (11–13,17). The present results support (but do not establish) this hypothesis. Although we cannot exclude that Pap-mediated contacts are probe dependent (i.e., not ordinarily made by PheB24 or ValA3), a direct interaction would rationalize the exquisite sensitivity of binding at each site to subtle modifications (such as TyrB24, AlaA3, ThrA3, and LeuA3) (9,10). A direct interaction is consistent with the structure and function of a truncated insulin analog lacking the COOH-terminal five residues of the B-chain (residues B26–B30) (magenta in Fig. 1A). In the crystal structure of this analog, ValA3 is exposed in an otherwise native-like conformation (18). When the new COOH-terminus is amidated, this analog is fully active (19). Conversely, tethering the COOH-terminal segment of the B-chain to the A-chain yields a native-like single-chain analog with essentially no biological activity (17). Furthermore, a conformational change in the B-chain would rationalize the low activity of a “chiral” analog in which the internal side chain of IleA2 (also shielded by TyrB26 and ProB28) is substituted by allo-isoleucine (10). This modification does not perturb the structure or stability of insulin but would alter its “hidden” functional surface (20).

In the decades since the crystal structure of insulin was elucidated in 1969 by D. Hodgkin et al. (3), the residues required for its function have been extensively investigated by mutagenesis and chemical modification (3,13). Interpretation of these results is incomplete, however, as such approaches do not generally distinguish between side chains that contact the receptor and those required to stabilize insulin’s active conformation. By exploiting site-specific photo cross-linking, the present studies strongly suggest that sites of clinical mutations (1,2) are in direct contact with the insulin receptor. A molecular understanding of such contacts, likely to emerge from a crystal structure of the hormone-receptor complex, may enable design of nonpeptide insulin agonists for the treatment of diabetes.

RESEARCH DESIGN AND METHODS

Insulin analogs were synthesized and purified as described (4,8). The relative receptor-binding affinities of para-amino-Phe analogs were determined by competitive displacement of 125I-insulin from a human placental membrane preparation, as previously described (10). Conversion of such analogs to Pap derivatives was effected as described (4). Lectin-purification of the insulin receptor overexpressed in cell line P3-A was performed by the procedure of Yoshimasa et al. (21). Photo cross-linking reactions were performed at high concentrations of hormone and receptor (ca. 200 nmol/l) to enable essentially complete binding of the PapA3 analog. Short-wave ultraviolet light (254 nm) generated from a Mineralight Lamp (Model UVG-54; UVP, Upland, CA) was used with an optimum exposure time of 20 s and a distance of 1 cm from the light source. Identification of photo cross-linked receptor domains utilized prior characterization of chymotryptic and tryptic sites (22,23). Western blots used Neutravidin (Pierce, IL) and a polyclonal anti-receptor antiserum that recognizes the NH2-terminal region of the α subunit (N-20; Santa Cruz Biotech, Santa Cruz, CA). For such mapping studies, analogs contained an amino-caproyl-biotin tag either at the α-amino group of residue B1 or, in the case of the second PapA3 derivative, the ε-amino group of d-lysine introduced in place of glycine at position A1. Domain-mapping studies are supported by SDS-PAGE analysis of cross-linked fragments following enzymatic deglycosylation as described (24).

FIG. 1.
  • Download figure
  • Open in new tab
  • Download powerpoint
FIG. 1.

Insulin structure and photo cross-linking. A: Ribbon model based on crystal structure (3) showing sites of clinical mutation (ValA3, PheB24, and PheB25; red) and terminal residues of A-chain (GlyA1 and AsnA21; blue). The A-chain is shown in silver and B-chain in gray or magenta (B26-B30). B and C: Photo cross-linking of Pap-modified insulin analogs (6 kDa) to ectodomain of insulin receptor (290 kDa α2β′2 tetramer; α subunit, 115 kDa; β′fragment, 30 kDa). Analysis of photo products (asterisks) by SDS-PAGE and Western blot using NeutrAvidin to detect biotin tag on insulin B-chain (NAv) or polyclonal antiserum to NH2-terminal peptide of the α-subunit (IRα-N; Santa Cruz Biotech). B: Top panel: Photo cross-linking via positions B25 (lane 4) and B24 (lane 8) analyzed after reduction by DTT. Lanes 1–3 and 5–7 indicate control reactions in which either the ectodomain was omitted (lanes 1, 2, 5, and 6) or samples not irradiated (lanes 1, 3, 5, and 7). Middle and bottom panels: Control blots demonstrating that equal amounts of ectodomain (middle panel; with DTT) and insulin (bottom panel; without DTT) were present in each reaction. C: Photo cross-linking via positions B25, A1, A3, and A21 (lanes 2, 6, 10, and 14, respectively) analyzed without reduction. D: Specificity of photo cross-linking is indicated by competition using native insulin (lanes 1–6) or IGF-1 (lanes 7–12). Protein concentrations in successive lanes are in each case 0-, 6-, 20-, 60-, 200-, and 600-fold greater than that of the photoreactive analog (PapB25). Efficiency of photo cross-linking is not affected by the addition of lysozyme or IgG as nonspecific competitors (not shown). In each experiment, the concentration of insulin analog and/or ectodomain was ca. 200 nmol/l in 50 mmol/l HEPES, 0.1% Triton X-100, and 110 mmol/l NaCl (pH 7.4).

FIG. 2.
  • Download figure
  • Open in new tab
  • Download powerpoint
FIG. 2.

Surface of insulin monomer (3). A: Space-filling representation (stereo view). Side chains of A3, B24, and B25 are shown in red and A21 in green. The A-chain is otherwise silver, and the B-chain is gray. B and C: Dot representation of restricted solvent-accessible surfaces (stereo pair) of ValA3 (B) and PheB24 (C). In a collection of crystallographic protomers (protein database identifiers 1APH, 1EV3, 1EV6, 1LPH, 1TRZ, and 4INS), respective solvent accessibilities of the A3 and B24 side chains are 14 ± 8 and 16 ± 2% relative to an extended GGXA tetrapeptide (4). Neighboring residues are as indicated; sulfur atoms are shown as yellow spheres. Although the native side chains are largely buried, the azido moieties at A3 and B24 are likely to protrude from the surface and so be exposed. Side chains of AsnA21 and PheB25 are exposed. PapA1 is predicted to be exposed based on the native-like crystal structure of TrpA1 insulin in which the indole ring projects near the B-chain COOH-terminus (15).

Acknowledgments

We thank G.D. Smith and C. Yip for kindly providing the receptor ectodomain and D.F. Steiner for a mammalian cell line overexpressing the human insulin receptor. S.H.N. was supported in part by the Diabetes Research & Training Center of the University of Chicago. This work was supported in part by grants from the National Institutes of Health to P.G.K. (DK56673) and M.A.W. (DK40949).

Footnotes

    • Accepted February 25, 2004.
    • Received December 11, 2003.
  • DIABETES

REFERENCES

  1. ↵
    Shoelson S, Haneda M, Blix P, Nanjo A, Sanke T, Inouye K, Steiner D, Rubenstein A, Tager H: Three mutant insulins in man. Nature302 :540 –543,1983
    OpenUrlCrossRefPubMedWeb of Science
  2. ↵
    Steiner DF, Tager HS, Chan SJ, Nanjo K, Sanke T, Rubenstein AH: Lessons learned from molecular biology of insulin-gene mutations. Diabetes Care13 :600 –609,1990
    OpenUrlAbstract/FREE Full Text
  3. ↵
    Baker EN, Blundell TL, Cutfield JF, Cutfield SM, Dodson EJ, Dodson GG, Hodgkin DM, Hubbard RE, Isaacs NW, Reynolds CD: The structure of 2Zn pig insulin crystals at 1.5 Å resolution. Phil Trans Royal Soc London Ser319 :369 –456,1988
    OpenUrl
  4. ↵
    Hua QX, Hu SQ, Frank BH, Jia W, Chu YC, Wang SH, Burke GT, Katsoyannis PG, Weiss MA: Mapping the functional surface of insulin by design: structure and function of a novel A-chain analogue. J Mol Biol264 :390 –403,1996
    OpenUrlCrossRefPubMedWeb of Science
  5. ↵
    Olsen HB, Ludvigsen S, Kaarsholm NC: Solution structure of an engineered insulin monomer at neutral pH. Biochemistry35 :8836 –8845,1996
    OpenUrlCrossRefPubMed
  6. ↵
    Nakagawa SH, Tager HS: Role of the phenylalanine B25 side chain in directing insulin interaction with its receptor: steric and conformational effects. J Biol Chem261 :7332 –7341,1986
    OpenUrlAbstract/FREE Full Text
  7. ↵
    Shoelson SE, Lee J, Lynch CS, Backer JM, Pilch PF: BpaB25 insulins: photoactivatable analogues that quantitatively cross-link, radiolabel, and activate the insulin receptor. J Biol Chem268 :4085 –4091,1993
    OpenUrlAbstract/FREE Full Text
  8. ↵
    Kurose T, Pashmforoush M, Yoshimasa Y, Carroll R, Schwartz GP, Burke GT, Katsoyannis PG, Steiner DF: Cross-linking of a B25 azidophenylalanine insulin derivative to the carboxyl-terminal region of the alpha-subunit of the insulin receptor: identification of a new insulin-binding domain in the insulin receptor. J Biol Chem269 :29190 –29197,1994
    OpenUrlAbstract/FREE Full Text
  9. ↵
    Mirmira RG, Tager HS: Role of the phenylalanine B24 side chain in directing insulin interaction with its receptor: importance of main chain conformation. J Biol Chem264 :6349 –6354,1989
    OpenUrlAbstract/FREE Full Text
  10. ↵
    Nakagawa SH, Tager HS: Importance of aliphatic side-chain structure at positions 2 and 3 of the insulin A chain in insulin-receptor interactions. Biochemistry31 :3204 –3214,1992
    OpenUrlCrossRefPubMed
  11. ↵
    Hua QX, Shoelson SE, Kochoyan M, Weiss MA: Receptor binding redefined by a structural switch in a mutant human insulin. Nature354 :238 –241,1991
    OpenUrlCrossRefPubMed
  12. Ludvigsen S, Olsen HB, Kaarsholm NC: A structural switch in a mutant insulin exposes key residues for receptor binding. J Mol Biol279 :1 –7,1998
    OpenUrlCrossRefPubMedWeb of Science
  13. ↵
    De Meyts P, Whittaker J: Structural biology of insulin and IGF-I receptors: implications for drug design. Nat Rev Drug Discovery1 :769 –783,2002
    OpenUrlCrossRefPubMedWeb of Science
  14. ↵
    Eberle AN, de Graan PNE: General principles for photoaffinity labeling of peptide hormone receptors. Methods Enzymol109 :129 –157,1985
    OpenUrlCrossRef
  15. ↵
    Wan ZL, Liang DC: Studies on the crystal structure of A1-(L-tryptophan) insulin at 2.1 Å resolution. Sci China Series B31 :1426 –1438,1988
    OpenUrl
  16. ↵
    Yip CC, Ottensmeyer P: Three-dimensional structural interactions of insulin and its receptor. J Biol Chem278 :27329 –27332,2003
    OpenUrlFREE Full Text
  17. ↵
    Derewenda U, Derewenda Z, Dodson EJ, Dodson GG, Bing X, Markussen J: X-ray analysis of the single chain B29–A1 peptide-linked insulin molecule: a completely inactive analogue. J Mol Biol220 :425 –433,1991
    OpenUrlCrossRefPubMedWeb of Science
  18. ↵
    Bi RC, Dauter Z, Dodson E, Dodson G, Giordano F, Reynolds C: Insulin structure as a modified and monomeric molecule. Biopolymers23 :391 –395,1984
    OpenUrlCrossRefWeb of Science
  19. ↵
    Cosmatos A, Ferderigos N, Katsoyannis PG: Chemical synthesis of [des(tetrapeptide B27–30), Tyr(NH2)26-B] and [des(pentapeptide B26–30), Phe(NH2)25-B] bovine insulins. Int J Protein Res14 :457 –471,1979
  20. ↵
    Xu B, Hua QX, Nakagawa SH, Jia W, Chu YC, Katsoyannis PG, Weiss MA: Chiral mutagensis of insulin’s hidden receptor-binding surface: structure of an allo-isoleucineA2 analogue. J Mol Biol316 :435 –441,2002
    OpenUrlCrossRefPubMedWeb of Science
  21. ↵
    Yoshimasa Y, Paul JI, Whittaker J, Steiner DF: Effects of amino acid replacements within the tetrabasic cleavage site on the processing of the human insulin receptor precursor expressed in Chinese hamster ovary cells. J Biol Chem265 :17230 –17237,1990
    OpenUrlAbstract/FREE Full Text
  22. ↵
    Boni-Schnetzler M, Scott W, Waugh SM, DiBella EE, Pilch PF: The insulin receptor: structural basis for high affinity ligand binding. J Biol Chem262 :8395 –8401,1987
    OpenUrlAbstract/FREE Full Text
  23. ↵
    Waugh SM, DiBella EE, Pilch PF: Isolation of a proteolytically derived domain of the insulin receptor containing the major site of cross-linking/binding. Biochemistry28 :3448 –3455,1989
    OpenUrlCrossRefPubMed
  24. ↵
    Xu B, Hu SQ, Chu YC, Huang K, Nakagawa SH, Whittaker J, Katosoyannis PG, Weiss MA: Diabetes-associated mutations in insulin: consecutive residues in the B chain contact distinct domains of the insulin receptor. Biochemistry In press
PreviousNext
Back to top

In this Issue

June 2004, 53(6)
  • Table of Contents
  • Index by Author
Sign up to receive current issue alerts
View Selected Citations (0)
Print
Download PDF
Article Alerts
Sign In to Email Alerts with your Email Address
Email Article

Thank you for your interest in spreading the word about Diabetes.

NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.

Enter multiple addresses on separate lines or separate them with commas.
Diabetes-Associated Mutations in Insulin Identify Invariant Receptor Contacts
(Your Name) has forwarded a page to you from Diabetes
(Your Name) thought you would like to see this page from the Diabetes web site.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
Diabetes-Associated Mutations in Insulin Identify Invariant Receptor Contacts
Bin Xu, Shi-Quan Hu, Ying-Chi Chu, Shuhua Wang, Run-ying Wang, Satoe H. Nakagawa, Panayotis G. Katsoyannis, Michael A. Weiss
Diabetes Jun 2004, 53 (6) 1599-1602; DOI: 10.2337/diabetes.53.6.1599

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Add to Selected Citations
Share

Diabetes-Associated Mutations in Insulin Identify Invariant Receptor Contacts
Bin Xu, Shi-Quan Hu, Ying-Chi Chu, Shuhua Wang, Run-ying Wang, Satoe H. Nakagawa, Panayotis G. Katsoyannis, Michael A. Weiss
Diabetes Jun 2004, 53 (6) 1599-1602; DOI: 10.2337/diabetes.53.6.1599
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • RESEARCH DESIGN AND METHODS
    • Acknowledgments
    • Footnotes
    • REFERENCES
  • Figures & Tables
  • Info & Metrics
  • PDF

Related Articles

Cited By...

More in this TOC Section

  • Genetic Variation at the ACE Gene Is Associated With Persistent Microalbuminuria and Severe Nephropathy in Type 1 Diabetes
  • Is Puberty an Accelerator of Type 1 Diabetes in IL6-174CC Females?
  • Association of the Diabetes Gene Calpain-10 With Subclinical Atherosclerosis
Show more Brief Genetics Report

Similar Articles

Navigate

  • Current Issue
  • Online Ahead of Print
  • Scientific Sessions Abstracts
  • Collections
  • Archives
  • Submit
  • Subscribe
  • Email Alerts
  • RSS Feeds

More Information

  • About the Journal
  • Instructions for Authors
  • Journal Policies
  • Reprints and Permissions
  • Advertising
  • Privacy Policy: ADA Journals
  • Copyright Notice/Public Access Policy
  • Contact Us

Other ADA Resources

  • Diabetes Care
  • Clinical Diabetes
  • Diabetes Spectrum
  • Scientific Sessions Abstracts
  • Standards of Medical Care in Diabetes
  • BMJ Open - Diabetes Research & Care
  • Professional Books
  • Diabetes Forecast

 

  • DiabetesJournals.org
  • Diabetes Core Update
  • ADA's DiabetesPro
  • ADA Member Directory
  • Diabetes.org

© 2021 by the American Diabetes Association. Diabetes Print ISSN: 0012-1797, Online ISSN: 1939-327X.