Tissue-type plasminogen activator protein

M. Mumenthaler, C. C. Hsu, and R. Pearlman, Feasibility study on spray-drying protein pharmaceuticals recombinant human growth hormone and tissue-type plasminogen activator, Pharm. Res, 11(1), 12 (1994). 

The heterogeneity of Lp(a) is related to ultrastructural motifs in the molecule, strongly resembling the so-called kringles found in both plasminogen and other plasma proteins, such as proteases of the coagulation system (tissue type plasminogen activator and prothrombin) (Fig. 2). 

Tincture of the dried seed, on agar plate at a concentration of 30 p,L/disc, was inactive on Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. Extract of 10 g plant material in 100 mL ethanol was used b Anticoagulation activity. Serpin BSZx (an inhibitor of trypsin and chemotrypsin) inhibited thrombin, plasma kallikrein, factor Vlla/tissue factor, and factor Xa at heparin-independent association rates. Only factor Xa turned a significant fraction of BSZx over as substrate. Activated protein C and leukocyte elastase were slowly inhibited by BSZx, whereas factor Xlla, urokinase and tissue type plasminogen activator, plasmin and pancreas kallikrein, and elastase were not or only weakly affected. Trypsin from Fusarium was not inhibited, while interaction with subtilisin Carlsberg and Novo was rapid, but most BSZx was cleaved as a substrate L... 

Bu, G., S. Williams, D.K. Strickland, and A.L. Schwartz. 1992. Low density lipoprotein receptor-related protein/alpha 2-macroglobulin receptor is an hepatic receptor for tissue-type plasminogen activator. Proc. Natl. Acad. Sci. USA... 

Yepes M, Sandkvist M, Moore EG, Bugge TH, Strickland DK, Lawrence DA (2003) Tissue-type plasminogen activator induces opening of the blood-brain barrier via the LDL receptor-related protein. J Clin Invest 112 1533-1540... 

Special applications of CIEF have focused on studies of glycoproteins, antibodies, and proteins in serum (e.g., hemoglobin variants [30,37,39,77-90], transferrin forms [21,31,36,80,82,90,91]), and cerebrospinal fluid [92], The gly-coforms of recombinant human tissue-type plasminogen activator [93-95] and... 

Mumenthaler M, Hsu CC, and Pearlman R. Feasibility study on Spray-drying Protein Pharmaceuticals Recombinant Human Growth Hormone and Tissue-type Plasminogen Activator. Pharm Res 1994 11(1) 12—20. 

Figure 10.13 shows the excellent performance of reversed-phase HPLC in biotechnological research. It represents the separation of the tryptic hydrolysate of the normal form and of a mutant of tissue-type plasminogen activator. This protein is built up of 527 amino acids and has a mass of approximately 67 000 Da. The mutant differs in a single amino acid, which leads to a deviating retention time of this specific fragment. 

Figure 10.13 Separation of the tryptic hydrolysate of tissue-type plasminogen activator [reproduced with permission from R.L. Garnick, N.J. Solli and P.A. Papa, Anal. Chem., 60,2546 (1988)]. Conditions stationary phase. Nova PakClS, 5 pm mobile phase Imlrnin" 50 mM sodium phosphate pH 2.8-acetonitrile, step gradient UV detector 210 nm. Top the normal protein with arginine at position 275 below the mutant with glutamic acid at position 275.

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