Angiostatin

Human

E.coli

Angiostatin

Angiostatin is an antiangiogenic 3845 kDa proteolytic fragment of Plasminogen, a 92100 kDa glycosylated blood zymogen that serves as the precursor for Plasmin (1). Plasminogen is produced primarily in the liver, but also in other tissues. Angiostatin circulates in the plasma, binds endothelial cells and myeloid cells, is present in platelet granules, and is excreted in the urine . Human Plasminogen contains an Nterminal activation peptide between amino acids (aa) 198, five characteristically folded kringle domains (aa 103561), and a peptidase S1 domain (aa 581808). Cleavage of the activation peptide produces mature Plasminogen, while further cleavage between Arg580 and Val581 by tPA (tissue plasminogen activator) produces the disulfidelinked twosubunit enzyme plasmin that dissolves fibrin clots. Angiostatin was first identified as consisting of kringles 14, a form called K14. Human Angiostatin (K14, aa 99459) shares 7980% aa sequence identity with mouse, rat, canine, feline, porcine and bovine K14. Other antiangiogenic forms include kringles 13 (K13), or 14 plus most of kringle 5 (K4.5). K4.5, which is reported to be the most active form, occurs in vivo by autoproteolysis of mature Plasminogen in the presence of either a sulfhydryl donor or cell surface actin, while matrix metalloproteins such as MMP3, 7, 9 and 19 can create multiple forms.

Approximately 29.7 KDa

VYLSECKTGN GKNYRGTMSK TKNGITCQKW SSTSPHRPRF SPATHPSEGL EENYCRNPDN DPQGPWCYTT DPEKRYDYCD ILECEEECMH CSGENYDGKI SKTMSGLECQ AWDSQSPHAH GYIPSKFPNK NLKKNYCRNP DRELRPWCFT TDPNKRWELC DIPRCTTPPP SSGPTYQCLK GTGENYRGNV AVTVSGHTCQ HWSAQTPHTH NRTPENFPCK NLDENYCRNP DGKRAPWCHT TNSQVRWEYC KIPSCDSSP

Sterile Filtered White lyophilized (freeze-dried) powder.

> 95% by SDS-PAGE and HPLC analyses.

The specific activity determined by an assay on anti-proliferation and anti-migration using endothelial cells in vitro and anti-angiogenesis in vivo is 5.5 × 105 IU/mg. Fully biologically active when compared to standard.

< 1 EU/μg of protein as determined by LAL method.

Lyophilized from a 0.2 μm filtered concentrated solution in 20 mM NaAc, pH 5.5, 4 % mannitol.

We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Reconstitute in sterile distilled water or aqueous buffer containing 0.1% BSA to a concentration of 0.1-1.0 mg/mL. Stock solutions should be apportioned into working aliquots and stored at ≤ -20°C. Further dilutions should be made in appropriate buffered solutions.

10, 100, 500μg