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PubMed2000

The cowpox virus serpin SPI-3 complexes with and inhibits urokinase-type and tissue-type plasminogen activators and plasmin.

The orthopoxvirus serpin SPI-3 is N-glycosylated and suppresses fusion between infected cells. Although SPI-3 contains motifs conserved in inhibitory serpins, no proteinase inhibition by SPI-3 has been demonstrated, and mutations within the serpin reactive center loop (RCL) do not affect the ability to regulate cell fusion. We demonstrate here that SPI-3 protein expressed by transcription/translation in vitro is able to form SDS-stable complexes with the serine proteinases plasmin, urokinase-type plasminogen activator (uPA), and tissue-type plasminogen activator (tPA), consistent with inhibitory activity of the serpin. Weaker complexes were noted with factor Xa and thrombin. Mutation of Arg-340/Ser-341 at the predicted P1/P1' sites within the RCL prevented the formation of complexes between SPI-3 and plasmin, uPA, or tPA, suggesting that the arginine at the P1 position was required for complex formation. SPI-3 protein lacking the N-terminal signal peptide was purified by means of an N-terminal His(10)-tag and gave complete inhibition in vitro of plasmin, uPA, and tPA and partial inhibition of factor Xa. SPI-3 is therefore a bifunctional protein that acts as a proteinase inhibitor and suppresses infected cell-cell fusion. As a proteinase inhibitor, SPI-3 has similar specificity to the leporipoxvirus SERP1 protein of myxoma virus, although the two serpins are less than 30% identical overall. The inhibition constants of SPI-3 for plasmin, uPA, and tPA were determined to be 0.64, 0.51, and 1.9 nM, respectively, very similar to the corresponding K(i) values of SERP1.

Acute-Phase Proteins↗
PubMed2000

[Cowpox and catpox infection. 2 Clinical case reports].

In patients with painful, possibly hemorrhagic vesicles or black crusts, especially on hands/fingers or face/neck with typical history (contact to cows or feral/cats) the possibility of a cowpox/catpox infection has to be considered. The clinical diagnosis can be confirmed with the electron microscopy; cytoplasmic inclusions may be detected histologically. Further useful diagnostic tools are the identification of the cowpox/catpox virus by PCR or in cell culture as well as serological tests to detect virus specific antibodies. We report the development of typical skin lesions of a cowpox/catpox infection in two female patients who had contact with cows or cats. Recent diagnostic and therapeutic approaches are also discussed.

Adult↗
PubMedDate not supplied

Cowpox and a cat.

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Adolescent↗