The binding of insulin-like growth factor II to the erythroleukemia cell line K562

1984 ◽  
Vol 4 (12) ◽  
pp. 1071-1077 ◽  
Author(s):  
Michael Tally ◽  
Tang Xin-Zhi ◽  
Gösta Enberg ◽  
Kerstin Hall
Keyword(s):  
Growth Factor ◽  
Cell Line ◽  
Specific Binding ◽  
Affinity Constant ◽  
Scatchard Analysis ◽  
Insulin Like Growth Factor ◽  
Factor Ii ◽  
Igf I ◽  
Erythroleukemia Cell ◽  
High Concentrations

The erythroleukemia cell line K562 was previously shown to have specific binding sites for insulin but not for insulin-like growth factor I (IGF-I). In this study the presence of specific receptors for insulin-like growth factor II (IGFqI) is established. Scatchard analysis of the competition curve for IGF-II disclosed a non-cooperative binding kinetic with a calculated affinity constant of 2.4 × 108 M−1 and a receptor number of 4.8 × 104 sites/cell. IGF-I displayed 10% crossreactivity over the IGF-II receptor but insulin did not crossreact at all. Instead insulin, present in high concentrations, enhanced the binding of IGF-II. The presence of IGF II but not IGF-I receptors makes the K562 cell line suitable for studying properties of the type-2 receptor.

Receptors for insulin-like growth factor-II in the growing tip of the deer antler

1993 ◽  
Vol 138 (2) ◽  
pp. 233-NP ◽  
Author(s):  
J. L. Elliott ◽  
J. M. Oldham ◽  
G. R. Ambler ◽  
P. C. Molan ◽  
G. S. G. Spencer ◽  
...  
Keyword(s):  
Growth Factor ◽  
Specific Binding ◽  
Cross Linking ◽  
Insulin Like Growth Factor ◽  
Deer Antler ◽  
Reducing Conditions ◽  
Factor Ii ◽  
Igf I ◽  
Igf Receptor

ABSTRACT Insulin-like growth factor-II (IGF-II) binding in the growing tip of the deer antler was examined using autoradiographical studies, radioreceptor assays and affinity cross-linking studies. Antler tips from red deer stags were removed 60 days after the commencement of growth, and cryogenically cut into sections. Sections were incubated with radiolabelled IGF-II, with or without an excess of competing unlabelled IGF-II and analysed autoradiographically. Radiolabelled IGF-II showed high specific binding in the reserve mesenchyme and perichondrium zones, which are tissues undergoing rapid differentiation and cell division in the antler. Binding to all other structural zones was low and significantly (P<0·001) less than binding to the reserve mesenchyme/perichondrium zones. Radioreceptor assays on antler microsomal membrane preparations revealed that the IGF-II binding was to a relatively homogeneous receptor population (Kd= 1·3 × 10−10 mol/l) with characteristics that were not entirely consistent with those normally attributed to the type 2 IGF receptor. Tracer binding was partly displaceable by IGF-I and insulin at concentrations above 10 nmol/l. However, affinity cross-linking studies revealed a single band migrating at 220 kDa under non-reducing conditions, indicative of the type 2 IGF receptor. These results indicate that, in antler tip tissues, IGF-II binds to sites which have different binding patterns and properties from receptors binding IGF-I. This may have functional significance as it appears that, whilst IGF-I has a role in matrix development of cartilage, IGF-II may have a role in the most rapidly differentiating and proliferating tissues of the antler. Journal of Endocrinology (1993) 138, 233–241

scholarly journals Insulin-like growth factor-I signaling is modified during chondrocyte differentiation

2004 ◽  
Vol 183 (3) ◽  
pp. 477-486 ◽  
Author(s):  
Chanika Phornphutkul ◽  
Ke-Ying Wu ◽  
Xu Yang ◽  
Qian Chen ◽  
Philip A Gruppuso
Keyword(s):  
Growth Factor ◽  
Cell Line ◽  
Chondrocyte Differentiation ◽  
Erk Pathway ◽  
Insulin Like Growth Factor ◽  
Erk Activation ◽  
Atdc5 Cell ◽  
Factor I ◽  
Proliferation And Differentiation ◽  
Igf I

Insulin-like growth factor-I (IGF-I) is a critical regulator of skeletal growth. While IGF-I has been shown to be a potent chondrocyte mitogen in vitro, its role in chondrocyte differentiation is less well characterized. We chose to study the action of IGF-I on an accepted model of chondrocyte differentiation, the ATDC5 cell line. Insulin concentrations sufficiently high to interact with the IGF-I receptor are routinely used to induce ATDC5 cells to differentiate. Therefore, we first examined the ability of IGF-I to promote chondrocyte differentiation at physiological concentrations. IGF-I could induce differentiation of these cells at concentrations below 10 nM. However, increasing IGF-I concentrations were less potent at inducing differentiation. We hypothesized that mitogenic effects of IGF-I might inhibit its differentiating effects. Indeed, the extracellular-signal-regulated kinase (ERK)-pathway inhibitor PD98059 inhibited ATDC5 cell DNA synthesis while enhancing differentiation. This suggested that the ability of IGF-I to promote both proliferation and differentiation might require that its signaling be modulated through the differentiation process. We therefore compared IGF-I-mediated ERK activation in proliferating and hypertrophic chondrocytes. IGF-I potently induced ERK activation in proliferating cells, but minimal ERK response was seen in hypertrophic cells. In contrast, IGF-I-mediated Akt activation was unchanged by differentiation, indicating intact upstream IGF-I receptor signaling. Similar findings were observed in the RCJ3.1C5.18 chondrogenic cell line and in primary chick chondrocytes. We conclude that IGF-I promotes both proliferation and differentiation of chondrocytes and that the differentiation effects of IGF-I may require uncoupling of signaling to the ERK pathway.

Measurement of Human Igf-II Using Sephacryl Extraction: A Rapid and Reliable Assay Method

1996 ◽  
Vol 33 (3) ◽  
pp. 201-208 ◽  
Author(s):  
Barbara H Mason ◽  
Michele A Tatnell ◽  
Ian M Holdaway
Keyword(s):  
Growth Factor ◽  
Binding Proteins ◽  
Complete Removal ◽  
Assay Method ◽  
Insulin Like Growth Factor ◽  
Serum Concentrations ◽  
Igf Binding Proteins ◽  
Factor Ii ◽  
Igf I ◽  
Igfbp 3

Measurement of insulin-like growth factor II (IGF-II) in human serum is complicated by the presence of IGF binding proteins and usually involves cumbersome extraction procedures followed by radioimmunoassay. We have utilized an extraction process developed for measuring insulin-like growth factor II in ovine serum using Sephacryl HR100, and have applied this to the extraction of human samples followed by radioimmunoassay for human IGF-II. The assay yielded 98% recovery of unlabelled IGF-II, parallelism between dilutions of eluate and the standard curve, complete removal of binding proteins and near-complete removal of IGF-I, and intra- and interassay coefficients of variation of 5% and 9%, respectively. The normal range for serum IGF-II in women was 490–1056 μg/L, and IGF-II levels were positively correlated with serum concentrations of insulin-like growth factor binding protein-3 (IGFBP-3) but not with IGF-I levels. Mean serum concentrations of IGF-II were reduced below normal in a number of hypopituitary patients and children with short stature and IGF-II concentrations in these subjects correlated positively with IGF-I and IGFBP-3. In acromegalic patients IGF-II levels were usually normal and were negatively correlated with IGF-I concentrations. From our experience with the above results the present assay appears particularly suitable for clinical measurements and research projects where high sample throughput is required.

Biosynthesis of 10 kDa and 7.5 kDa insulin-like growth factor II in a human rhabdomyosarcoma cell line

1993 ◽  
Vol 93 (1) ◽  
pp. 87-95 ◽  
Author(s):  
Finn C. Nielsen ◽  
Gisela Haselbacher ◽  
Jan Christiansen ◽  
Mats Lake ◽  
Mette Grønborg ◽  
...  
Keyword(s):  
Growth Factor ◽  
Cell Line ◽  
Insulin Like Growth Factor ◽  
Rhabdomyosarcoma Cell Line ◽  
Factor Ii ◽  
Rhabdomyosarcoma Cell

Retinoic acid regulates insulin-like growth factor II expression in a neuroblastoma cell line.

Endocrinology ◽  
1992 ◽  
Vol 130 (6) ◽  
pp. 3669-3676 ◽  
Author(s):  
K Matsumoto ◽  
C Gaetano ◽  
W H Daughaday ◽  
C J Thiele
Keyword(s):  
Retinoic Acid ◽  
Growth Factor ◽  
Cell Line ◽  
Neuroblastoma Cell ◽  
Neuroblastoma Cell Line ◽  
Insulin Like Growth Factor ◽  
Factor Ii

A monoclonal antibody to human insulin-like growth factor-I: characterization, use in radioimmunoassay and effect on the biological activities of the growth factor

1989 ◽  
Vol 2 (3) ◽  
pp. 201-206 ◽  
Author(s):  
D. J. Morrell ◽  
H. Dadi ◽  
J. More ◽  
A. M. Taylor ◽  
A. Dabestani ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Growth Factor ◽  
Binding Site ◽  
Biological Activities ◽  
Rabbit Antiserum ◽  
Affinity Constant ◽  
Insulin Like Growth Factor ◽  
Factor I ◽  
Igf I ◽  
Growth Factor I

ABSTRACT A monoclonal antibody (BPL-M23) to insulin-like growth factor-I (IGF-I) was obtained following immunization of BALB/c mice with human IGF-I conjugated to ovalbumin. The affinity constant of BPL-M23 for IGF-I was 10·5 litres/nmol and the cross-reactivities of IGF-II, multiplication-stimulating activity III-2 and insulin were 08, 003 and less than 0·0001 % respectively. Porcine, bovine, ovine and rabbit sera, but not rat or mouse sera, showed substantial reactivity with the antibody. Comparison of radioimmunoassay analyses of 54 human serum samples from normal subjects and acromegalic and GH-deficient patients using BPL-M23 and a polyclonal rabbit antiserum (R557A) to human IGF-I showed a high correlation, indicating the usefulness of the monoclonal antibody in radioimmunoassay. Monoclonal antibody BPL-M23 was capable of abolishing the sulphation, mitogenic and insulin-like activities of IGF-I in in-vitro bioassays, suggesting that these activities may rely upon the same receptor-binding site which is near to the antibody-binding site.

Hypoglycemia in a dog with a leiomyoma of the gastric wall producing an insulin-like growth factor II-like peptide

1995 ◽  
Vol 132 (6) ◽  
pp. 744-750 ◽  
Author(s):  
Andrea Boari ◽  
Antonina Barreca ◽  
Gilberto E Bestetti ◽  
Francesco Minuto ◽  
Massimiliano Venturoli
Keyword(s):  
Growth Factor ◽  
Tumor Tissue ◽  
Tissue Concentration ◽  
Molecular Form ◽  
Islet Cell Tumor ◽  
Gastric Wall ◽  
Insulin Like Growth Factor ◽  
Factor Ii ◽  
Igf I ◽  
Tumor Hypoglycemia

Boari A, Barreca A, Bestetti GE, Minuto F, Venturoli M. Hypoglycemia in a dog with a leiomyoma of the gastric wall producing an insulin-like growth factor II-like peptide. Eur J Endocrinol 1995;132:744–50. ISSN 0804–4643 A 12-year-old mixed-breed male dog was referred to the Clinica Medica Veterinaria of Bologna University for recurrent episodes of seizures due to hypoglycemia with abnormally low plasma insulin levels (18 pmol/l), Resection of a large leiomyoma (780 g) of the gastric wall resulted in a permanent resolution of the hypoglycemic episodes. Insulin-like growth factors I and II (IGF-I and -II) were measured by RIA in serum before and after surgery and in tumor tissue. Results were compared to the serum concentration of 54 normal and to the tissue concentration observed in eight non-hypoglycemic dog gastric wall extracts. Before surgery, circulating immunoreactive IGF-I was 0.92 nmol/l, which is significantly lower than the control values (16.92 ± 8.44 nmol/l, range 3.53–35.03), while IGF-II was 152 nmol/l, which is significantly higher than the control values (42.21 ± 3.75. range 31.99–50.74). After surgery, IGF-I increased to 6.80 nmol/l while IGF-II decreased to 45.52 nmol/l, Tumor tissue IGF-II concentration was higher than normal (5.66 nmol/kg tissue as compared to a range in normal gastric wall tissue of 1.14–3.72 nmol/kg), while IGF-I was 0.08 nmol/kg tissue, which is close to the lowest normal value (range in controls, 0.08–1.18 nmol/kg). Partial characterization of IGF-II immunoreactivity extracted from tissue evidenced a molecular weight similar to that of mature IGF-II thus excluding that peptide released by the tumor is a precursor molecule. In agreement with these data, at variance with samples of normal dog gastric wall, IGF-II immunostaining was positive and in situ hybridization evidenced the expression of IGF-II mRNA in tumor tissue specimen. Evaluation of the molecular distribution of the IGFs in the circulation evidenced that IGF-II immunoreactivity was predominantly in the 3 5–65 kD region and barely detectable in the other regions. These results show that in dog, non-islet cell tumor hypoglycemia, as demonstrated in humans, can be ascribed to overproduction of IGF-II circulating in a molecular form that can more easily cross the capillary wall, thus exerting its insulin-like effects on target tissues. Andrea Boari, Istituto di Clinica Medical Veterinaria di Bologna, Via Tolara di Sopra, 40064 Ozzano Emilia (BO), Italy

Rat somatotroph insulin-like growth factor-II (IGF-II) signaling: role of the IGF-I receptor.

Endocrinology ◽  
1992 ◽  
Vol 131 (5) ◽  
pp. 2147-2153 ◽  
Author(s):  
M M Weber ◽  
S Melmed ◽  
J Rosenbloom ◽  
H Yamasaki ◽  
D Prager
Keyword(s):  
Growth Factor ◽  
Insulin Like Growth Factor ◽  
Factor Ii ◽  
Igf I
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