Home LiteratureArticle Details
PMID: 4170389 Published · ppublish English Journal Article

The nature of human serum insulin-like activity (ILA)P Characterization of ILA in serum and serum fractions obtained by acid-ethanol extraction and adsorption chromatography.

The Journal of clinical investigation ·Vol. 47 ·No. 2 ·1968-02-00 ·Pages 301-20

Poffenbarger PL, Ensinck JW, Hepp DK, Williams RH

Abstract

Studies were undertaken in an attempt to clarify the apparent heterogeneous nature of human serum insulin-like activity. Methods of preparative zone electrophoresis on Pevikon, acid-ethanol extraction of trichloroacetic acid serum protein precipitates, adsorption chromatography on DEAE-cellulose and Dowex 50, gel filtration chromatography, and insulin antiserum immunoreactivity were used. The results establish the presence of a substance in serum with in vitro biological properties similar to insuln but with different physicochemical properties. The major portion of serum ILA measured by bioassay techniques can be attributed to the effects of this substance. Whereas the in vitro biological effects of this substance on muscle and adipose cells were similar to those of crystalline insulin, the substance is distinguished from insulin by: (1) the failure of insulin antiserum to inhibit its in vitro biological effect; (2) a slower electrophoretic mobility (in the gamma-beta globulin zone); and (3) a larger molecular weight, between 40,000 and 50,000 in these studies. It is similar to insulin since both are soluble in acid-ethanol. The results further indicate that previously described insulin-like activity in gamma-beta globulin preparations, the major portion of total serum insulin activity described in acid-ethanol extracts of serum, "bound" insulin, "atypical" insulin, and antibody nonsuppressible insulin-like activity bioassayed in diluted serum are all one and the same substance.

MeSH Terms
Adipose Tissue/drug effects,metabolism Animals Beta-Globulins/analysis Biological Assay Blood Protein Electrophoresis Carbon Isotopes Cellulose Chromatography, Gel Chromatography, Ion Exchange Diaphragm Ethanol Glucose/metabolism Hot Temperature Humans Hydrogen-Ion Concentration Insulin/blood Insulin Antibodies/pharmacology Molecular Weight Rats Trichloroacetic Acid gamma-Globulins/analysis
Chemicals
Beta-Globulins Carbon Isotopes Insulin Insulin Antibodies gamma-Globulins Ethanol Trichloroacetic Acid Cellulose Glucose
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Poffenbarger P L
Ensinck J W
Hepp D K
Williams R H
References (58)
58 references, click to expand
  1. THE INSULIN AND THE ZINC CONTENT OF NORMAL AND DIABETIC PANCREAS.
    J Clin Invest. 1938 Nov;17(6):725-8 PMID: 16694619
  2. Measurement of small quantities of insulin-like activity with rat adipose tissue. III. Persistence of serum insulin-like activity after pancreatectomy.
    J Clin Invest. 1962 Sep;41:1699-707 PMID: 13916770
  3. Studies of insulin antagonism. I. An artifactual antagonist to insulin and plasma nonsuppressible insulin-like activity occurring in preparation of "albumin".
    Diabetes. 1967 May;16(5):289-301 PMID: 6022555
  4. Serum insulin-like activity in hypophysectomized and depancreatized (Houssay) dogs.
    Diabetes. 1965 Oct;14(10):658-62 PMID: 5830628
  5. ANTIBODY-SUPPRESSIBLE AND NONSUPPRESSIBLE INSULIN-LIKE ACTIVITIES IN HUMAN SERUM AND THEIR PHYSIOLOGIC SIGNIFICANCE. AN INSULIN ASSAY WITH ADIPOSE TISSUE OF INCREASED PRECISION AND SPECIFICITY.
    J Clin Invest. 1963 Nov;42:1816-34 PMID: 14083170
  6. A COLORIMETRIC METHOD FOR SERUM CHYMOTRYPSIN DETERMINATION.
    Can J Biochem. 1965 Feb;43:135-41 PMID: 14325965
  7. The presence of two forms of insulin in normal human serum.
    Biochemistry. 1963 Mar-Apr;2:286-9 PMID: 13976977
  8. RELEASE OF INSULIN-LIKE ACTIVITY FROM SERUM OF NORMAL SUBJECTS AND DIABETIC PATIENTS.
    Metabolism. 1965 Feb;14:110-4 PMID: 14261395
  9. An assay for insulin-like activity using rat adipose tissue.
    Lancet. 1958 Jul 12;2(7037):76-7 PMID: 13564766
  10. Insulin-like activity of dilute human serum assayed by an isolated adipose cell method.
    Diabetes. 1965 Oct;14(10):643-9 PMID: 5890522
  11. Studies on the state of insulin in blood. Examination of splenic, portal and peripheral blood serum of diabetic and nondiabetic subjects for "free" insulin and insulin complexes.
    N Engl J Med. 1962 Aug 2;267:218-22 PMID: 13861796
  12. Studies on the state of insulin in blood: dissociation of purified human blood insulin complex(es) by incubation with adipose tissue extracts in vitro.
    Endocrinology. 1961 Jan;68:36-42 PMID: 13683766
  13. Separation of human plasma protein concentrate with insulin activity.
    Science. 1958 Mar 14;127(3298):593-4 PMID: 13529025
  14. Insulin treatment and increased UDPG-glycogen transglucosylase activity in muscle.
    Arch Biochem Biophys. 1961 Sep;94:436-42 PMID: 13781408
  15. Assay of insulin in blood.
    Physiol Rev. 1960 Apr;40:219-44 PMID: 13840871
  16. Studies on the state of insulin in blood: the role of glucose in the in vivo dissociation of insulin complexes.
    Endocrinology. 1961 Jul;69:163-9 PMID: 13683765
  17. [INSULIN ACTIVITY OF PROTEIN FRACTIONS OF NORMAL HUMAN SERUM].
    Pathol Biol (Paris). 1964 Feb;12:148-60 PMID: 14137102
  18. Effect of pancreatic proteins on insulin assay systems.
    Diabetes. 1966 May;15(5):342-5 PMID: 5936462
  19. The insulin activity of protein fractions of normal human serum.
    J Endocrinol. 1958 Nov;17(4):387-95 PMID: 13611151
  20. INSULIN ACTIVITY OF NORMAL PLASMA AND PLASMA ACIDETHANOL EXTRACTS.
    Lancet. 1964 Jan 11;1(7324):74-6 PMID: 14075541
  21. PURIFICATION AND STUDY OF THE PRINCIPAL PRODUCTS CONTAINED IN PREPARATIONS OF RADIOINSULIN.
    Metabolism. 1965 Feb;14:115-21 PMID: 14261396
  22. A NEW METHOD FOR RECOVERY OF INSULIN ACTIVITY FROM BLOOD-SERUM.
    Lancet. 1963 Sep 28;2(7309):656-8 PMID: 14052023
  23. Antigenicity of insulin: diabetes induced by specific antibodies.
    Biochem J. 1955 Feb;59(2):179-85 PMID: 14351177
  24. STIMULATION BY HYPEROSMOLARITY OF GLUCOSE METABOLISM IN RAT ADIPOSE TISSUE AND DIAPHRAGM IN VITRO.
    J Biol Chem. 1965 Jun;240:2277-83 PMID: 14304826
  25. The estimation of glycogen with the anthrone reagent.
    Arch Biochem. 1950 Jan;25(1):191-200 PMID: 15401229
  26. STUDIES EMPLOYING A NEW METHOD FOR THE RECOVERY OF BIOLOGICALLY ACTIVE INSULIN FROM ACID ALCOHOLIC EXTRACTS OF PANCREAS AND BLOOD SERUM.
    Diabetes. 1963 Sep-Oct;12:448-53 PMID: 14067745
  27. Some current controversies in diabetes research.
    Diabetes. 1965 Sep;14(9):549-72 PMID: 5318828
  28. THE INSULIN-LIKE ACTIVITY IN SERUM DETERMINED BY THE RAT EPIDIDYMAL FAT METHOD. V. THE DISTRIBUTION OF INSULIN-LIKE ACTIVITY IN ELECTROPHORETICALLY-SEPARATED SERUM PROTEIN FRACTIONS FROM NORMAL FASTING SUBJECTS AND THE EFFECT OF INGESTION OF GLUCOSE.
    Acta Med Scand. 1964 Apr;175:401-8 PMID: 14149646
  29. Non-suppressible insulin-like activity of human serum. II. Biological properties of plasma extracts with non-suppressible insulin-like activity.
    Biochim Biophys Acta. 1966 Jun 29;121(2):360-74 PMID: 5962523
  30. Non-suppressible insulin-like activity of human serum. I. Physicochemical properties, extraction and partial purification.
    Biochim Biophys Acta. 1966 Jun 29;121(2):349-59 PMID: 5962522
  31. ANABOLIC RESPONSES OF DIAPHRAGM MUSCLE TO INSULIN AND TO OTHER PANCREATIC PROTEINS.
    Proc Soc Exp Biol Med. 1964 Jul;116:669-71 PMID: 14194629
  32. The Vallance-Owen ("synalbumin") insulin antagonist. Reproducibility and reliability of results in nondiabetic and diabetic humans.
    Diabetes. 1966 Mar;15(3):149-59 PMID: 5909494
  33. Effect of insulin on the formation of glycogen by the mouse diaphragm in the presence and absence of glucose. A preliminary report.
    Diabetes. 1966 Jul;15(7):492-6 PMID: 5945211
  34. Preparation of iodine-131 labelled human growth hormone of high specific activity.
    Nature. 1962 May 5;194:495-6 PMID: 14450081
  35. Biological activity of blood insulin complexes examined by rat diaphragm tissue assay.
    Proc Soc Exp Biol Med. 1960 Jul;104:411-3 PMID: 13830286
  36. The assay of insulin with anti-insulin and mouse diaphragm.
    Can J Biochem Physiol. 1961 Apr;39:695-712 PMID: 13783129
  37. Further immunological studies on the form of circulating insulin.
    J Endocrinol. 1962 Jun;24:263-77 PMID: 14496556
  38. Insulin-like activity of human plasma constituents. II. Biologic assay of human plasma fractions for insulin-like activity.
    Metabolism. 1956 Jan;5(1):44-50 PMID: 13279549
  39. A COMPARISON OF INSULIN-LIKE ACTIVITY CONTAINED IN THE SERUM PROTEINS OF NORMAL SUBJECTS AND DIABETIC PATIENTS.
    Diabetes. 1965 Feb;14:71-6 PMID: 14256054
  40. Regeneration of insulin activity from the separated and inactive A and B chains.
    Nature. 1960 Nov 26;188:721-4 PMID: 13723142
  41. Changes in levels of 'atypical' circulating insulin after infusing 'typical' insulin through the liver.
    J Endocrinol. 1963 Mar;26:1-10 PMID: 13991273
  42. Responses of obese-hyperglycemic mice and normal mice to "bound" and crystalline insulin.
    Metabolism. 1966 Apr;15(4):325-9 PMID: 5905290
  43. Electrophoretic separation of insulin activity of normal plasma and plasma with added insulin-I 131.
    Metabolism. 1959 Jan;8(1):39-48 PMID: 13622235
  44. THE "TYPICAL" AND "ATYPICAL" FORMS OF SERUM INSULIN.
    Diabetes. 1963 Jul-Aug;12:339-48 PMID: 14087498
  45. Studies on the state of insulin in blood: materials and methods for the estimation of "free" and "bound" insulin-like activity in serum.
    Endocrinology. 1962 Jan;70:95-8 PMID: 13861798
  46. Insulin-like effects of serum albumin and globulin fractions on glucose uptake by rat epididymal adipose tissue.
    Diabetes. 1959 Nov-Dec;8:438-41 PMID: 13798330
  47. Improved separation of serum proteins in paper electrophoresis: a new electrophoresis buffer.
    Scand J Clin Lab Invest. 1957;9(4):338-41 PMID: 13495359
  48. STUDIES ON THE STATE OF INSULIN IN BLOOD: PROPERTIES OF CIRCULATING "FREE" AND "BOUND" INSULIN.
    Endocrinology. 1965 Apr;76:709-21 PMID: 14289193
  49. Studies on the state of insulin in blood: the state and transport of insulin in blood.
    Endocrinology. 1961 Jan;68:7-16 PMID: 13683767
  50. PROPERTIES OF HUMAN "ALBUMIN". A METABOLIC STUDY OF ALBUMIN EXTRACTS FROM NORMAL AND DIABETIC PLASMA.
    Diabetes. 1963 Sep-Oct;12:406-13 PMID: 14067738
  51. Plasma insulin in health and disease.
    Am J Med. 1961 Dec;31:874-81 PMID: 13868361
  52. Abnormalities of serum insulin-like activity in liver-disease.
    Lancet. 1962 Dec 22;2(7269):1287-90 PMID: 13991272
  53. Protein measurement with the Folin phenol reagent.
    J Biol Chem. 1951 Nov;193(1):265-75 PMID: 14907713
  54. Studies on the state of insulin in blood: "free" insulin and insulin complexes in human sera and and their in vitro biological properties.
    Endocrinology. 1961 Jul;69:46-54 PMID: 13683764
  55. EFFECTS OF TRYPSIN AND CHYMOTRYPSIN ON BLOOD GLUCOSE IN VIVO AND GLUCOSE UPTAKE IN VITRO.
    Proc Soc Exp Biol Med. 1965 Jul;119:866-9 PMID: 14329022
  56. The effect of insulin antiserum on the insulin activity of serum and protein fractions of serum.
    J Endocrinol. 1959 Dec;19:221-31 PMID: 13837173
  57. EFFECT OF ANAEROBIOSIS AND CELL POISONS ON GLUCOSE UPTAKE OF HEMIDIAPHRAGMS AND EPIDIDYMAL FAT PADS IN VITRO.
    Diabetes. 1965 Mar;14:128-31 PMID: 14262219
  58. Influence of oxygenation and deoxygenation on distribution of insulin activity among blood-plasma-protein fractions.
    Lancet. 1960 Dec 10;2(7163):1269-71 PMID: 13703590
Article Info
Journal
The Journal of clinical investigation
Abbr.
J Clin Invest
ISSN
0021-9738
DOI
10.1172/JCI105726
Published
1968-02-00
Pages
301-20
Language
English
Region
United States
NLM ID
7802877
PMCID
PMC297172
Subset
IM
External Links
PubMed source DOI: 10.1172/JCI105726 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2