Plasminogen mutants activated by thrombin. Potential thrombus-selective thrombolytic agents.

Dawson, Keith M.; Cook, A; Devine, Janet; Edwards, Richard M.; Hunter, M. G.; Raper, Robert H.; Roberts, Gordon C. K.

doi:10.1016/s0021-9258(17)33962-5

Cited by 20 publications

(2 citation statements)

References 26 publications

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“…During a mutagenesis project on plasminogen it was noted that a single-residue substitution variant [Plg(R719E)j, containing glutamic acid at residue 719 in the protease domain, was less readily activated than the wild-type molecule upon the addition of SK (Figure 2). This effect is unlikely to be a result of the expression system per se, as recombinant wildtype plasminogen and recombinant thrombin-activatable plasminogen T1 (Dawson et al, 1994) each possess SK activation activities dissimilar to that of Plg(R719E) and similar to that of natural human plasminogen, using the same assay regime (data not shown). The effect is likely to be due either to significant structural perturbation within the activa- Molecular Elution Time (Te, minutes)…”

Section: Discussionmentioning

confidence: 95%

Substitution of Arginine 719 for Glutamic Acid in Human Plasminogen Substantially Reduces Its Affinity for Streptokinase

Dawson¹,

Marshall²,

Raper³

et al. 1994

Biochemistry

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In isolation human plasminogen possesses no enzymatic activity, yet upon formation of an equimolar complex with the bacterial protein streptokinase, it acquires a plasminogen activator function. The region(s) of plasminogen and of streptokinase which mediate complex formation has (have) not been previously published. Here it is reported that a single-residue substitution (Arg719-->Glu) in the serine protease domain of full-length Glu-plasminogen substantially reduces its affinity for streptokinase. The plasminogen variant displays no other significant differences from the wild-type molecule with respect to activation by two-chain urokinase-type plasminogen activator, recognition by monoclonal antibodies, or ability to undergo conformational change. It is concluded that Arg719 in human plasminogen is an important determinant of the streptokinase binding site, although further sites are likely to contribute both to the affinity of plasminogen for streptokinase and to mechanisms by which the active site is formed within the complex.

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Section: Discussionmentioning

confidence: 95%

Substitution of Arginine 719 for Glutamic Acid in Human Plasminogen Substantially Reduces Its Affinity for Streptokinase

Dawson¹,

Marshall²,

Raper³

et al. 1994

Biochemistry

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“…Furthermore, our nanoformulation is designed for systemic thrombosis prevention in contrast to their hydrogel platform, which is better suited for highly localized anticoagulation applications (e.g., surface coating of blood contacting medical devices). 8 Engineering approaches have also been used to design bioresponsive thrombolytics ; antithrombotic therapies designed to dissolve existing clots rather than prevent their formation ; including recombinant proteins 47,50 or polymeric microparticles 51 activated by proteolytic or biophysical triggers associated with thrombosis that decrease bleeding and expand therapeutic windows. Looking forward, several clinical applications warrant further investigation with our nanoSTATs.…”

Section: Discussionmentioning

confidence: 99%

Self-Titrating Anticoagulant Nanocomplexes That Restore Homeostatic Regulation of the Coagulation Cascade

Lin¹,

Consul³

et al. 2014

ACS Nano

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Antithrombotic therapy is a critical portion of the treatment regime for a number of life-threatening conditions, including cardiovascular disease, stroke, and cancer; yet, proper clinical management of anticoagulation remains a challenge because existing agents increase the propensity for bleeding in patients. Here, we describe the development of a bioresponsive peptide–polysaccharide nanocomplex that utilizes a negative feedback mechanism to self-titrate the release of anticoagulant in response to varying levels of coagulation activity. This nanoscale self-titrating activatable therapeutic, or nanoSTAT, consists of a cationic thrombin-cleavable peptide and heparin, an anionic polysaccharide and widely used clinical anticoagulant. Under nonthrombotic conditions, nanoSTATs circulate inactively, neither releasing anticoagulant nor significantly prolonging bleeding time. However, in response to life-threatening pulmonary embolism, nanoSTATs locally release their drug payload and prevent thrombosis. This autonomous negative feedback regulator may improve antithrombotic therapy by increasing the therapeutic window and decreasing the bleeding risk of anticoagulants.

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Expression of the transcription factor GADD153 is an indicator of apoptosis for recombinant chinese hamster ovary (CHO) cells

Murphy

Woods²,

Dickson

2001

Biotech & Bioengineering

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Loss of cell viability, through engagement of apoptotic cell death, represents a limitation to maintenance of high levels of productivity of recombinant animal cells in culture. The ability to monitor the status of recombinant cells, and to define indicators of their "well-being," would present a valuable approach to permit a rational intervention at appropriate times during culture. Growth arrest and DNA damage gene 153 (GADD153) is a member of the CCAAT/enhancer-binding protein (C/EBP) family of transcription factors and has been associated with apoptosis. We have examined the expression of GADD153 in conditions associated with apoptosis of recombinant CHO cells in batch culture. GADD153 expression is very low in CHO cells growing in the exponential phase of batch culture but is activated as cells enter the decline phase. Depletion of nutrients (glucose or glutamine) causes activation of GADD153 expression as does the imposition of endoplasmic reticulum stress. In all cases, there is a good relationship between the extent of apoptosis that occurs in response to each stress and the degree of GADD153 expression. In addition, nutrient refeeding or reversal of stress produces a concomitant decrease in expression of GADD153 and the susceptibility to apoptosis. Thus, GADD153 appears to offer a valid indicator of apoptosis and illustrates the potential for definition of monitors of cellular status related to the likelihood of apoptosis of cell populations.

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Plasminogen mutants activated by thrombin. Potential thrombus-selective thrombolytic agents.

Cited by 20 publications

References 26 publications

Substitution of Arginine 719 for Glutamic Acid in Human Plasminogen Substantially Reduces Its Affinity for Streptokinase

Substitution of Arginine 719 for Glutamic Acid in Human Plasminogen Substantially Reduces Its Affinity for Streptokinase

Self-Titrating Anticoagulant Nanocomplexes That Restore Homeostatic Regulation of the Coagulation Cascade

Expression of the transcription factor GADD153 is an indicator of apoptosis for recombinant chinese hamster ovary (CHO) cells

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