Insulin-like growth factors I and II in healthy man.

1989 ◽  
Vol 121 (6) ◽  
pp. 753-758 ◽  
Author(s):  
Hans-Peter Guler ◽  
Jürgen Zapf ◽  
Christoph Schmid ◽  
E. Rudolf Froesch

Abstract. IGF-I and -II share specific serum carrier proteins which elute on neutral Sephadex G-200 gel permeation chromatography at apparent molecular masses of 50 and 200 kD. The half-lives of free and carrier protein-bound 125I-IGF-I and -II were determined after bolus injections of the tracers into two normal adults. Labelled IGF-I and -II migrated first with the 50-kD and later with the 200-kD complex. In these complexes their apparent half-lives were 20–30 min and 12–15 h, respectively. The apparent half-life of free 125I-IGF-I and -II was 10–12 min. In a second set of experiments, recombinant human insulin-like growth factor I was infused during 6 days in two healthy adults at a dose of 20 μg · kg−1 · h−1 (corresponding to around 30 mg/day). Serum obtained before and during the infusion was subjected to neutral Sephadex G-200 gel permeation chromatography and fractions were pooled according to the apparent molecular masses at which the carrier protein complexes elute. IGF-I and -II in these pools were determined by RIA. Before the IGF-I infusion, 92 and 272 μg/l of IGF-I and -II were found in the 200-kD complex, 45 and 91 μg/l in the 50-kD complex, and 15 and 5 μg/l were present in the free form. Corresponding figures during the IGF-I infusion were 389 and 18 μg/l for the 200-Kd complex, 201 and 54 μg/l for the 50-kD complex, and 80 and < 1 μg/l for free IGF-I and -II. Using the half-lives of the tracer studies and the levels of the different molecular weight forms of IGF in serum, the production rates for IGF-I and -II were calculated to be 10 mg and 13 mg per day.

1986 ◽  
Vol 32 (2) ◽  
pp. 363-367 ◽  
Author(s):  
W Flapper ◽  
P J van den Oetelaar ◽  
C P Breed ◽  
J Steenbergen ◽  
H J Hoenders

Abstract Human sera were subjected to analytical ultracentrifugation and "high-pressure" gel-permeation chromatography on a system of combined TSK Gel G5000 PW and G3000 SW columns. The chromatographic method produced remarkably superior resolution of the proteins, especially those exceeding 100 000 Da. We calculated the molecular masses of eluted fractions on the basis of their detection by low-angle laser light scattering and their differential refractive index. We discuss the results in relation to the clinical data.


1985 ◽  
Vol 110 (1) ◽  
pp. 140-144 ◽  
Author(s):  
Rose-Marie Schimpff ◽  
Mauro Bozzola ◽  
Jurgen Zapf ◽  
Anne-Marie Repellin ◽  
I. Einschenk

Abstract. Growth-promoting activity measured as [3H] thymidine incorporation into lectin-activated lymphocytes was determined simultaneously with radioimmunoassayable IGF I and IGF II in cord and capillary blood collected from human neonates 30 min and 24 h after birth. All the parameters studied in cord blood were lower than in normal adults. During the early postnatal period, IGF I decreased and IGF II remained unchanged, but thymidine activity increased above the normal adult level. The differences between the values found in cord blood and in capillary blood collected within the first ½ h after birth agree with a production of growth factors by the infant. All these growth factors may play a role in foetal growth, as suggested by their correlation with birth weight. Finally, factors other than IGFs contributing to thymidine activity may play a role in neonatal growth since they are higher in newborns than in adults.


2009 ◽  
Vol 51 (7) ◽  
pp. 785-790 ◽  
Author(s):  
A. I. Kuzaev ◽  
I. P. Kim ◽  
D. P. Kiryukhin ◽  
V. M. Buznik

The intramolecularly crosslinked polystyrene molecules, prepared as in part I of this series, were examined by the techniques of light scattering, osmometry, viscometry, and gel permeation chromatography. Their molecular masses and molecular mass distribution remained constant over all the stages of the reaction radii of gyration were determined and the results compared with those predicted in part II of this series.


1991 ◽  
Vol 37 (12) ◽  
pp. 2043-2047 ◽  
Author(s):  
László Góth

Abstract The different electrophoretic mobilities of erythrocyte and serum catalase (EC 1.11.1.6) were confirmed and the causes responsible for their differences were examined. The presence of a catalase-binding protein in serum that could form a complex with erythrocyte catalase was excluded by incubating serum proteins with erythrocyte catalase. No new unequivocal catalase bands representing a catalase-binding protein were detected. The erythrocyte and serum catalase proved to be charge isoforms: their molecular masses, estimated by gel permeation chromatography or polyacrylamide gel electrophoresis in a nondenaturing system, were very similar, whereas their electrophoretic mobilities were different. Assay of serum catalase by gel permeation and hydrophobic chromatography yielded a product with the same electrophoretic mobility as that of erythrocyte catalase. Different dilution of erythrocyte catalase with human sera led to a gradual decrease of its mobility, 20-fold or greater dilution yielding the same results as for serum catalase. Similarly, when serum catalase was diluted 20-fold or more with 60 mmol/L phosphate buffer, it migrated similarly to erythrocyte catalase. I detected no effect of dialyzable serum ligands, NADPH, or protection of SH groups on the electrophoretic mobility of either catalase isoform. I conclude that formation of charge isoforms of catalase is caused by a reversible, conformational modification due to matrix effect of serum.


1996 ◽  
Vol 42 (4) ◽  
pp. 307-315 ◽  
Author(s):  
Lesya A. Trachuk ◽  
Lyudmila P. Revina ◽  
Tatyana M. Shemyakina ◽  
Galina G. Chestukhina ◽  
Valentin M. Stepanov

Five chitinases were isolated from culture filtrates of Bacillus licheniformis B-6839 R and S variants by combination of hydrophobic, ion-exchange, and gel permeation chromatography. The enzymes had molecular masses of 66, 62, 53, 49, and 42 kDa. The chitinases revealed two activity optima against colloidal chitin at pH 4.5–5.5 and 9.0–9.5 and they were rather stable at pH 4.0–9.5. The temperature optimum of activity was 90 °C for the 62-kDa chitinase and 70 °C for the other enzymes. The 66-, 53-, and 42-kDa chitinases showed pronounced similarities in their N-terminal sequences and apparently belonged to the same group, which might be related to Bacillus circulans chitinase A1. The 49- and 62-kDa enzymes did not reveal structural similarities with other chitinases produced by the studied B. licheniformis strain. No relationship was found with the 89- and 76-kDa chitinases isolated earlier from B. licheniformis X-7u.Key words: Bacillus licheniformis, chitinase, multiplicity.


1987 ◽  
Vol 241 (3) ◽  
pp. 745-750 ◽  
Author(s):  
H J Cornell ◽  
G Enberg ◽  
A C Herington

Ion-exchange chromatography of serum on DEAE-Sephadex A-50 using a stepwise NaCl gradient showed that complexes enriched with insulin-like growth factors I and II (IGF-I and IGF-II) could be preferentially eluted. A fraction eluted with 0.075 M-NaCl preferentially contained immunoreactive IGF-I with peak levels appearing in fractions of Mr approx. 110,000. The IGF-I-binding protein complex itself had low bioactivity as measured in a non-suppressible insulin-like (NSILA) bioassay. On conversion to free IGF-I by gel-permeation chromatography on Sephadex G-75 in 1% formic acid, however, the IGF-I did express its intrinsic NSILA bioactivity. In contrast, an IGF-II-enriched complex was eluted from the DEAE-Sephadex with 0.15 M-NaCl. Practically all of the recovered NSILA of the original serum was present in this fraction, in the Mr range 70,000-300,000 with a peak of 150,000. Chromatography on Sephadex G-75 in 1% formic acid separated this high-Mr NSILA into low-Mr (less than 15000) IGF-II and high-Mr acid-stable NSILA-P. The high-Mr IGF-II complex bound to concanavalin A-Sepharose, suggesting that it was a glycoprotein. The results confirm previous reports that a large portion of the NSILA of whole serum can be accounted for by a biologically active acid-dissociable complex. These data show for the first time that this active complex consists of an IGF-II-preferring binding protein. In direct contrast, the IGF-I-preferring complex does not express NSILA bioactivity until the IGF-I is liberated through acidification. The presence of a metabolically active IGF-II complex in serum raises questions as to its possible biological role in the adult.


1995 ◽  
Vol 147 (3) ◽  
pp. 507-516 ◽  
Author(s):  
K M Hua ◽  
S C Hodgkinson ◽  
J J Bass

Abstract Plasma levels of IGFs-I and -II were measured in 4-month-old ewe lambs (n=20) and 2-year-old ewes (n=16), which were well fed (n=18) or fasted (n=18) for 3 days. Half of each nutrition group was given daily (0900 h) injections of bovine GH (bGH, 0·1 mg/kg body weight per day) for 3 days. Blood samples were collected immediately before the GH injection every morning. Plasma IGFs were extracted by acid gel permeation chromatography using a Waters Protein Pak 125 column, fitted to a Pharmacia fast protein liquid chromatography system, then freeze-dried, reconstituted (at pH 7·4) and estimated by RIA. At the end of the experiment, IGF-I levels in plasma were increased (P<0·01) by exogenous bGH in both fed ewes and lambs but not in the fasted animals; plasma IGF-I levels were depressed by fasting (P<0·01) at all ages. IGF-I levels were also found to be significantly higher (P<0·01) in ewes than lambs. In contrast, plasma IGF-II concentrations were depressed (P=0·02) by administration of bGH in all groups and elevated in the ewes (P<0·05) by fasting. However, the lambs showed no significant changes in IGF-II with fasting. The IGF-II levels were significantly higher (P<0·001) in lambs than ewes. Results from the present study demonstrate that GH administration stimulated an increase in plasma IGF-I and induced a decrease in plasma IGF-II. On the other hand, fasting depressed plasma IGF-I and elevated plasma IGF-II in the sheep. A significant GH/nutrition interaction for IGF-I (P<0·01), but not for IGF-II, and a significant nutrition/age interaction for IGF-II (P<0·01), but not for IGF-I, in the present study suggest that GH has a greater stimulating effect on plasma levels of IGF-I in the fed rather than fasted sheep and that nutrition has a greater influence on plasma levels of IGF-II in the older rather than younger animals, indicating that plasma IGFs-I and -II are differentially regulated by nutrition, GH and developmental stage in postnatal sheep. Journal of Endocrinology (1995) 147, 507–516


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