Serpin Polymerization Is Prevented by a Hydrogen Bond Network That Is Centered on His-334 and Stabilized by Glycerol

Zhou, Aiwu; Stein, Penelope E.; Huntington, James A.; Carrell, Robin W.

doi:10.1074/jbc.m211663200

Cited by 62 publications

(88 citation statements)

References 49 publications

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“…This mutation disrupts an important hydrogen bond network within the shutter domain and mimics the effects of the Mmalton and Siiyama mutations. 29 Both of these substitutions generate a mutant of ␣ 1 -antitrypsin that favors polymer formation more readily than the wild type protein. The mutants were polymerized by heating at 0.5 mg/mL and 50°C (His334Ala ␣ 1 -antitrypsin) or 55°C (Glu342Ala ␣ 1 -antitrypsin) for 30 minutes, and the resulting polymers were assessed by nondenaturing PAGE and Western blot analysis.…”

Section: Resultsmentioning

confidence: 99%

“…28 Recombinant Glu342Ala and His334Ala ␣ 1 -antitrypsin were expressed and purified as described previously. 29 Polymers of these mutants were prepared by heating at 0.5 mg/mL and 50°C (His334Ala ␣ 1 -antitrypsin) or 55°C (Glu342Ala Western Blot Analysis. Nondenaturing polyacrylamide gel electrophoresis (PAGE) was performed in a Mini-PROTEAN II electrophoresis cell (Bio-Rad Laboratories, Hercules, CA) on 7.5% wt/vol acrylamide gels as described previously.…”

Section: Methodsmentioning

confidence: 99%

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Differential detection of PAS-positive inclusions formed by the Z, Siiyama, and Mmalton variants of α1-antitrypsin

et al. 2004

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Differential detection of PAS-positive inclusions formed by the Z, Siiyama, and Mmalton variants of α1-antitrypsin

et al. 2004

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“…The His334Asp mutation is in the shutter region where it will disrupt a critical hydrogen bond network that controls opening of b-sheet A. 29 Both mutations result in polymers that are recognized by the 2C1 antibody suggesting that they share the same structure. Given the homology to the highly polymerogenic His338Arg variant of neuroserpin, it is likely that neuroserpin mutants form polymers with a similar structure to those formed by a 1 -antitrypsin.…”

Section: Discussionmentioning

confidence: 99%

A Novel Monoclonal Antibody to Characterize Pathogenic Polymers in Liver Disease Associated with α1-Antitrypsin Deficiency†

et al. 2010

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Alpha 1 -antitrypsin is the most abundant circulating protease inhibitor. The severe Z deficiency allele (Glu342Lys) causes the protein to undergo a conformational transition and form ordered polymers that are retained within hepatocytes. This causes neonatal hepatitis, cirrhosis, and hepatocellular carcinoma. We have developed a conformation-specific monoclonal antibody (2C1) that recognizes the pathological polymers formed by a 1 -antitrypsin. This antibody was used to characterize the Z variant and a novel shutter domain mutant (His334Asp; a 1 -antitrypsin King's) identified in a 6-week-old boy who presented with prolonged jaundice. His334Asp a 1 -antitrypsin rapidly forms polymers that accumulate within the endoplasmic reticulum and show delayed secretion when compared to the wild-type M a 1 -antitrypsin. The 2C1 antibody recognizes polymers formed by Z and His334Asp a 1 -antitrypsin despite the mutations directing their effects on different parts of the protein. This antibody also recognized polymers formed by the Siiyama (Ser53Phe) and Brescia (Gly225Arg) mutants, which also mediate their effects on the shutter region of a 1 -antitrypsin. Conclusion: Z and shutter domain mutants of a 1 -antitrypsin form polymers with a shared epitope and so are likely to have a similar structure. (HEPATOLOGY 2010;52:1078-1088 T he serpinopathies are conformational diseases characterized by the polymerization and intracellular retention of members of the serine protease inhibitor or serpin superfamily of proteins.1 The best known is a 1 -antitrypsin deficiency, with the most common severe deficiency allele being the Z mutation (Glu342Lys). This mutation results in the retention of ordered polymers of a 1 -antitrypsin as periodic acid Schiff positive inclusion bodies within the endoplasmic reticulum (ER) of hepatocytes.2 These inclusions predispose the individual homozygous for the Z variant of the a 1 -antitrypsin protease inhibitor (PI*Z) to neonatal hepatitis, cirrhosis, and rarely, hepatocellular carcinoma.3 Deficiency of circulating a 1 -antitrypsin results in early onset panlobular emphysema.

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“…Of particular importance in this respect is the P8 residue (Zhou et al, 2003), which is a threonine in most inhibitory serpins (Thr359 in ShSPI).…”

Section: Shutter Regionmentioning

confidence: 99%

Three-dimensional structure of a schistosome serpin revealing an unusual configuration of the helical subdomain

Granzin¹,

Huang²,

Topbaş³

et al. 2012

Acta Crystallogr D Biol Crystallogr

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Parasitic organisms are constantly challenged by the defence mechanisms of their respective hosts, which often depend on serine protease activities. Consequently, protease inhibitors such as those belonging to the serpin superfamily have emerged as protective elements that support the survival of the parasites. This report describes the crystal structure of ShSPI, a serpin from the trematode Schistosoma haematobium. The protein is exposed on the surface of invading cercaria as well as of adult worms, suggesting its involvement in the parasite–host interaction. While generally conforming to the well established serpin fold, the structure reveals several distinctive features, mostly concerning the helical subdomain of the protein. It is proposed that these peculiarities are related to the unique biological properties of a small serpin subfamily which is conserved among pathogenic schistosomes.

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Serpin Polymerization Is Prevented by a Hydrogen Bond Network That Is Centered on His-334 and Stabilized by Glycerol

Cited by 62 publications

References 49 publications

Differential detection of PAS-positive inclusions formed by the Z, Siiyama, and Mmalton variants of α1-antitrypsin

Differential detection of PAS-positive inclusions formed by the Z, Siiyama, and Mmalton variants of α1-antitrypsin

A Novel Monoclonal Antibody to Characterize Pathogenic Polymers in Liver Disease Associated with α1-Antitrypsin Deficiency†

Three-dimensional structure of a schistosome serpin revealing an unusual configuration of the helical subdomain

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