The amplified mouse squamous cell carcinoma antigen gene locus contains a serpin (Serpinb3b) that inhibits both papain-like cysteine and trypsin-like serine proteinases

Askew, David J.; Askew, Yuko S.; Kato, Yukari; Luke, Cliff J.; Pak, Stephen C.; Brὂmme, Dieter; Silverman, Gary A.

doi:10.1016/j.ygeno.2004.01.014

Cited by 17 publications

(20 citation statements)

References 31 publications

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“…Sequences were analyzed using MacVector 6.0 (Accelrys Inc., Princeton, NJ). Serpinb11 was amplified and cloned from a mouse lung cDNA library (Clontech) as described (18,20).…”

Section: Methodsmentioning

confidence: 99%

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SERPINB11 Is a New Noninhibitory Intracellular Serpin

Askew

Çataltepe

Kumar

et al. 2007

Journal of Biological Chemistry

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SERPINB11, the last of 13 human clade B serpins to be described, gave rise to seven different isoforms. One cDNA contained a premature termination codon, two contained splice variants, and four contained full-length open reading frames punctuated by eight single nucleotide polymorphisms (SNPs). The SNPs encoded amino acid variants located within the serpin scaffold but not the reactive site loop (RSL). Although the mouse orthologue, Serpinb11, could inhibit trypsin-like peptidases, SERPINB11 showed no inhibitory activity. To determine whether the human RSL targeted a different class of peptidases or the serpin scaffold was unable to support inhibitory activity, we synthesized chimeric human and mouse proteins, in which the RSLs had been swapped. The human RSL served as a trypsin inhibitor when supported by mouse scaffold sequences. Conversely, the mouse RSL on the human scaffold showed no inhibitory activity. These findings suggested that variant residues in the SERPINB11 scaffold impaired serpin function. SDS-PAGE analysis supported this notion as RSL-cleaved SERPINB11 was unable to undergo the stressed-to-relaxed transition typical of inhibitory type serpins. Mutagenesis studies supported this hypothesis, since the reversion of amino acid sequences in helices D and I to those conserved in other clade B serpins partially restored the ability of SERPINB11 to form covalent complexes with trypsin. Taken together, these findings suggested that SER-PINB11 SNPs encoded amino acids in the scaffold that impaired RSL mobility, and HapMap data showed that the majority of genomes in different human populations harbored these noninhibitory SERPINB11 alleles. Like several other serpin superfamily members, SERPINB11 has lost inhibitory activity and may have evolved a noninhibitory function.Serpins are a superfamily of serine and cysteine peptidase inhibitors that contain ϳ1500 family members and are found in all domains of life (Eukarya, Eubacteria, and Archaea) as well as many Poxviridae (reviewed in Refs. 1-4). Unlike canonical inhibitors, inhibitory serpins employ a unique suicide substrate-like inhibitory mechanism to neutralize their target peptidases. The surface-exposed reactive site loop (RSL) 4 serves as a pseudosubstrate and binds to the active site of the peptidase. Upon cleavage of the RSL, the metastable serpin molecule undergoes a major conformational rearrangement and traps the covalently attached peptidase in a distorted, inactive form (5). A small proportion of serpins are noninhibitory and aid in diverse functions, such as hormone transport (e.g. SERPINA7/ thyroxine binding globulin) and protein folding (e.g. SER-PINH1/HSP47) (6, 7).Serpins are classified into 17 clades based on phylogenetic relationships (8). So far, 36 human serpins from nine clades (A-I) have been identified (2). The majority of serpins are plasma proteins that serve as critical regulators of important physiological processes, such as blood coagulation, fibrinolysis, and inflammation. In contrast, clade B serpins exist predominantly, but...

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“…Sequences were analyzed using MacVector 6.0 (Accelrys Inc., Princeton, NJ). Serpinb11 was amplified and cloned from a mouse lung cDNA library (Clontech) as described (18,20).…”

Section: Methodsmentioning

confidence: 99%

“…Similarly, first-strand cDNA samples from adult mouse tissues and mouse embryos were screened for Serpinb11 transcripts by PCR using specific primers (forward 1, 5Ј-GGGTA-AAAGTGCAGTTGTGAATATG-3Ј; reverse 1, 5Ј-GGAGAA-GCAAACTTGCCAGC-3Ј) (18,20). These primers amplified a 554-bp product.…”

Section: Matrix-assisted Laser Desorption Ionization Mass Spectroscopmentioning

confidence: 99%

SERPINB11 Is a New Noninhibitory Intracellular Serpin

Askew

Çataltepe

Kumar

et al. 2007

Journal of Biological Chemistry

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“…These molecules function as intracellular inhibitors of cysteine and serine proteases with distinct cross-class specificity, protecting cells from aberrant proteolysis by a general mechanism, using a C-terminal reactive site loop as bait and acting as a suicide substrate (44,45). Among these is Serpinb3a, the product of which specifically inhibits Ctsg (46,47) and which, along with Serpinb3b and Serpinb3c, forms part of the SCCA locus in mice (referred to herein as serpinb3a/b/c) (48). Using qRT-PCR, we confirmed upregulation of serpinb3a/b/c in the lungs of IFN-γ-blocked Nos2 -/-mice compared with control mice at day 28 p.i.…”

Section: Figurementioning

confidence: 99%

Serine protease activity contributes to control of Mycobacterium tuberculosis in hypoxic lung granulomas in mice

Reece¹,

Loddenkemper²,

Askew³

et al. 2010

J. Clin. Invest.

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The hallmark of human Mycobacterium tuberculosis infection is the presence of lung granulomas. Lung granulomas can have different phenotypes, with caseous necrosis and hypoxia present within these structures during active tuberculosis. Production of NO by the inducible host enzyme NOS2 is a key antimycobacterial defense mechanism that requires oxygen as a substrate; it is therefore likely to perform inefficiently in hypoxic regions of granulomas in which M. tuberculosis persists. Here we have used Nos2 -/-mice to investigate host-protective mechanisms within hypoxic granulomas and identified a role for host serine proteases in hypoxic granulomas in determining outcome of disease. Nos2 -/-mice reproduced human-like granulomas in the lung when infected with M. tuberculosis in the ear dermis. The granulomas were hypoxic and contained large amounts of the serine protease cathepsin G and clade B serine protease inhibitors (serpins). Extrinsic inhibition of serine protease activity in vivo resulted in distorted granuloma structure, extensive hypoxia, and increased bacterial growth in this model. These data suggest that serine protease activity acts as a protective mechanism within hypoxic regions of lung granulomas and present a potential new strategy for the treatment of tuberculosis.

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“…The SCCA locus in the mouse contains 4 genes of which Serpinb3a, also known as SQN-5 and SCCA-2, is clearly related to both SCCA-1 and SCCA-2 in humans. Mouse serpinb3a is both a cysteine-and trypsin-like serine proteinase inhibitor (9), and it is expressed in tissues in which human SCCA-1 and SCCA-2 are also expressed. Thus, Serpinb3a, which is referred to as SCCA-2 in this paper, is the mouse ortholog of human SCCA-1 and SCCA-2 (6).…”

Section: Uteroglobin (Ug)mentioning

confidence: 99%

Uteroglobin Suppresses SCCA Gene Expression Associated with Allergic Asthma

Ray

Choi

Zhang

et al. 2005

Journal of Biological Chemistry

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Uteroglobin (UG), the founding member of the Secretoglobin superfamily, is a potent anti-inflammatory protein constitutively expressed at a high level in the airway epithelia of all mammals. We previously reported that the lungs of UG-knock-out (UG-KO) mice express elevated levels of Th2 cytokines (e.g. interleukin (IL)-4 and IL-13), which are augmented by allergen sensitization and challenge leading to exaggerated airway inflammation. Notably, these responses are suppressed by recombinant UG treatment (Mandal, A. K., Zhang, Z., Ray, R., Choi, M. S., Chowdhury, B., Pattabiraman, N., and Mukherjee, A. B. (2004) J. Exp. Med. 199, 1317-1330). Recent reports indicate that human orthologs of murine squamous cell carcinoma antigen-2 (SCCA-2/serpinb3a), a serine proteaseinhibitor, are overexpressed in the airways of asthmatic patients. We report here that compared with wild type littermates, UG-KO mouse lungs express markedly elevated levels of SCCA-2 mRNA and protein, which are augmented by allergen-challenge. Most importantly, these effects are abrogated by recombinant UG treatment. We further demonstrate that treatment of cultured human bronchial epithelial cells with IL-4 or IL-13 stimulates phosphorylation of STAT-1 and STAT-6 leading to SCCA-1 (SERPINB3) and SCCA-2 (SERPINB4) gene expression. We propose that: (i) IL-4-and IL-13-stimulated SCCA gene expression is mediated via STAT-1 and STAT-6 activation, and (ii) by suppressing the production, and most likely by interfering with the signaling of these cytokines, UG inhibits SCCA gene expression associated with airway inflammation in asthma. Uteroglobin (UG)1 is a steroid-inducible, homodimeric, multifunctional secreted protein with potent anti-inflammatory and immunomodulatory properties (reviewed in Ref. 1). The founding member of the secretoglobin superfamily of proteins (2), UG, is constitutively expressed at a high level in the pulmonary mucosal epithelial cells of virtually all mammals including mice. We previously reported that compared with wild type (WT) littermates, the lungs of the UG-knock-out (UG-KO) mice express markedly higher levels of Th2 cytokines and manifest exaggerated airway inflammatory response to allergens marked by elevated levels of eosinophil infiltration (3, 4). All of these are characteristically found in human airway inflammatory diseases such as bronchial asthma in which sensitivity to allergens play a critical pathogenic role.Recently, it has been reported that the expression of squamous cell carcinoma antigens (SCCA), cysteine, and chymotrypsin proteinase inhibitors of the ovalbumin/serpin family (reviewed in Ref. 5; Ref. 6) is markedly elevated in the airway epithelia of patients with bronchial asthma (7). Consistent with these findings, it has also been reported that SCCA protein targets a potent allergen and an extrinsic cysteine proteinase in house-dust mites, which is thought to play a pathogenic role in allergic asthma (8), although the physiological role(s) of SCCA is yet to be defined.In humans, SCCA-1 and SCCA-2 genes enc...

show abstract

The amplified mouse squamous cell carcinoma antigen gene locus contains a serpin (Serpinb3b) that inhibits both papain-like cysteine and trypsin-like serine proteinases

Cited by 17 publications

References 31 publications

SERPINB11 Is a New Noninhibitory Intracellular Serpin

SERPINB11 Is a New Noninhibitory Intracellular Serpin

Serine protease activity contributes to control of Mycobacterium tuberculosis in hypoxic lung granulomas in mice

Uteroglobin Suppresses SCCA Gene Expression Associated with Allergic Asthma

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