Osteoprotegerin Human Pichia Pastoris

Introduction to the Molecule

Osteoprotegerin (OPG) or osteoclastogenesis inhibitory factor (OCIF) is a secretory glycoprotein belonging to TNF receptor superfamily. OPG consists of 401 amino acid residues, it has a molecular weight of 60 kDa as a monomer and 120 kDa as a disulfide-linked dimer and is produced in different tissues, e.g. bone, skin, liver, stomach, intestine and lung. Osteoprotegerin inhibits the recruitment, proliferation and activation of osteoclasts. Osteoclast formation activity may be monitored principally by determination of concentration ratio of osteoprotegerin ligand (OPGL)/OPG. Alteration of this ratio may be the cause of bone loss in many imbalances in bone metabolism such as osteoporosis, osteopetrosis, hypercalcemia, metastatic osteolytic lesions and rheumatic bone degradation.

Research topic

Bone and cartilage metabolism

Description

Recombinant OPG produced in yeast contains 412 amino acid residues, including 180 residues from mature OPG (a.a 22–201) and 232 residues from the Fc protein of human IgG1, and has a calculated molecular mass of 46.5 kDa. As a result of glycosylation, the recombinant Osteoprotegrin migrates as a 49 kDa protein in SDS-PAGE under reducing conditions. The OPG is purified by proprietary chromatographic techniques.

Amino Acid Sequence

Source

Pichia Pastoris

Purity

Greater than 90.0% as determined by

(a) Analysis by RP-HPLC.

(b) Analysis by SDS-PAGE.

Biological Activity

Determined by its ability to neutralize the stimulation of U937 cells treated with 10ng/ml of soluble RANKL (sRANKL).

Formulation

OPG was lyophilized from a 0.2?m filtered concentrated (0.5mg/ml) solution in PBS, pH= 7.4.

Storage, Stability/Shelf Life

It is recommended to reconstitute the lyophilized Osteoprotegerin in sterile 18 M?-cm H2O not less than 100 µg/ml, which can then be further diluted to other aqueous solutions.

Physical Appearance

Sterile Filtered White lyophilized (freeze-dried) powder.

References

• Bonnelye E, Aubin JE. Osteoprotegerin and its ligand: A new paradigm for regulation of osteoclastogenesis and bone resorption. Medscape Womens Health. 2000 Mar;5 (2):5
• Boyle WJ, Lacey DL, Xu W, Tan HL, Scully S, Capparelli C, Tarpley J, Morony S, Dunstan CR, Sarosi I, Bucay N, Simonet WS. osteoprotegerin-deficient mice develop early onset osteoporosis and arterial calcification. Genes Dev. 1998 May 1;12 (9):1260-8
• Dollerup J, Gormsen LC, Schmitz O, Mogensen CE, Rasmussen LM, Nielsen S, Nellemann B. Simvastatin reduces plasma osteoprotegerin in type 2 diabetic patients with microalbuminuria. Diabetes Care. 2007 Dec;30 (12):3122-4
• Hofbauer LC. Osteoprotegerin ligand and osteoprotegerin: novel implications for osteoclast biology and bone metabolism. Eur J Endocrinol. 1999 Sep;141 (3):195-210
• Karan M, Lam A, Mangan S, McCann M, Golledge J. Osteoprotegerin and osteopontin are expressed at high concentrations within symptomatic carotid atherosclerosis. Stroke. 2004 Jul;35 (7):1636-41
• Kerschan-Schindl K, Fureder W, Mitterbauer M, Kudlacek S, Grampp S, Bieglmayer C, Fialka-Moser V, Pietschmann P, Kalhs P. Bone metabolism in patients more than five years after bone marrow transplantation. Bone Marrow Transplant. 2004 Sep;34 (6):491-6
• Lee R, Derby P, Tarpley J, Van G, Sander S, Campbell P, Pattison W, Hill D, Hughes TM, Renshaw-Gegg L, Bucay N, Davy E, Sullivan J, Trail G, Tan HL, Colombero A, Elliott R, DeRose M, Shimamoto G, Boone T, Bennett L, Wooden S, Nguyen HQ, Luthy R, Chang MS, Kelley M, Dunstan CR, Lacey DL, Simonet WS, Boyle WJ. Osteoprotegerin: a novel secreted protein involved in the regulation of bone density. Cell. 1997 Apr 18;89 (2):309-19
• Moran CS, McCann M, Karan M, Norman P, Ketheesan N, Golledge J. Association of osteoprotegerin with human abdominal aortic aneurysm progression. Circulation. 2005 Jun 14;111 (23):3119-25
• Morse LR, Nguyen HP, Jain N, Williams S, Tun CG, Battaglino RA, Stashenko P, Garshick E. Age and motor score predict osteoprotegerin level in chronic spinal cord injury. J Musculoskelet Neuronal Inter. 2008 Jan-Mar;8 (1):50-7
• Ostensen M, Leistad L, Sundan A, Seidel C, Borset M, Feuerherm AJ, Faxvaag A. Elevated levels of osteoprotegerin (OPG) and hepatocyte growth factor (HGF) in rheumatoid arthritis. Scand J Rheumatol. 2001;30 (4):229-34
• Secchiero P, Corallini F, Pandolfi A, Consoli A, Candido R, Fabris B, Celeghini C, Capitani S, Zauli G. An increased osteoprotegerin serum release characterizes the early onset of diabetes mellitus and may contribute to endothelial cell dysfunction. Am J Pathol. 2006 Dec;169 (6):2236-44
• Sultan A, Piot C, Mariano-Goulart D, Avignon A, Thuan Dit Dieudonne JF, Cristol JP, Dupuy AM. Osteoprotegerin: a novel independent marker for silent myocardial ischemia in asymptomatic diabetic patients. Diabetes Care. 2007 Nov;30 (11):2934-9
• Ueland T, Bollerslev J, Godang K, Muller F, Froland SS, Aukrust P. Increased serum osteoprotegerin in disorders characterized by persistent immune activation or glucocorticoid excess--possible role in bone homeostasis. Eur J Endocrinol. 2001 Dec;145 (6):685-90
• Vilim V, Vytasek R, Olejarova M, Machacek S, Gatterova J, Prochazka B, Kraus VB, Pavelka K. Serum cartilage oligomeric matrix protein reflects the presence of clinically diagnosed synovitis in patients with knee osteoarthritis. Osteoarthritis Cartilage. 2001 Oct;9 (7):612-8
• Yamada Y, Higashio K, Ikeda K, Harada A, Goto M, Kobayashi F, Washida N, Tsuda E, Yano K. Immunological characterization of circulating osteoprotegerin/osteoclastogenesis inhibitory factor: increased serum concentrations in postmenopausal women with osteoporosis. J Bone Miner Res. 1999 Apr;14 (4):518-27

By research topic:

• Bone and cartilage metabolism

By molecule:

• Osteoprotegerin