Contribution of a Single-Turn α-Helix to the Conformational Stability and Activity of the Alkaline Proteinase Inhibitor ofPseudomonasaeruginosa

Biochemistry ◽  
2005 ◽  
Vol 44 (7) ◽  
pp. 2469-2477 ◽  
Author(s):  
Robert D. Gray ◽  
John O. Trent
Keyword(s):  
Proteinase Inhibitor ◽  
Conformational Stability ◽  
Alkaline Proteinase ◽  
Single Turn ◽  
Α Helix

Molecular basis for defective secretion of the Z variant of human alpha-1-proteinase inhibitor: secretion of variants having altered potential for salt bridge formation between amino acids 290 and 342

1989 ◽  
Vol 9 (4) ◽  
pp. 1406-1414
Author(s):  
A A McCracken ◽  
K B Kruse ◽  
J L Brown
Keyword(s):  
Amino Acid ◽  
Proteinase Inhibitor ◽  
Conformational Stability ◽  
Point Mutations ◽  
Salt Bridge ◽  
Site Directed Mutagenesis ◽  
Wild Type ◽  
Functional Importance ◽  
Bridge Formation ◽  
Cos Cells

Human alpha-1-proteinase inhibitor (A1PI) deficiency, associated with the Z-variant A1PI (A1PI/Z) gene, results from defective secretion of the inhibitor from the liver. The A1PI/Z gene exhibits two point mutations which specify amino acid substitutions, Val-213 to Ala and Glu-342 to Lys. The functional importance of these substitutions in A1PI deficiency was investigated by studying the secretion of A1PI synthesized in COS cells transfected with A1PI genes altered by site-directed mutagenesis. This model system correctly duplicates the secretion defect seen in individuals homozygous for the A1PI/Z allele and shows that the substitution of Lys for Glu-342 alone causes defective secretion of A1PI. The substitution of Lys for Glu-342 eliminates the possibility for a salt bridge between residues 342 and 290, which may decrease the conformational stability of the molecule and thus account for the secretion defect. However, when we removed the potential to form a salt bridge from the wild-type inhibitor by changing Lys-290 to Glu (A1PI/SB-290Glu), secretion was not reduced to the 19% of normal level seen for A1PI/Z-342Lys; in fact, 75% of normal secretion was observed. When the potential for salt bridge formation was returned to A1PI/Z-342Lys by changing Lys-290 to Glu, only 46% of normal secretion was seen. These data indicate that the amino acid substitution at position 342, rather than the potential to form the 290-342 salt bridge, is the critical alteration leading to the defect in A1PI secretion.

scholarly journals Alkaline Proteinase Inhibitor ofPseudomonas aeruginosa

2000 ◽  
Vol 275 (28) ◽  
pp. 21002-21009 ◽  
Author(s):  
Rhona E. Feltzer ◽  
Robert D. Gray ◽  
William L. Dean ◽  
William M. Pierce
Keyword(s):  
Proteinase Inhibitor ◽  
Alkaline Proteinase

NMR structure note: alkaline proteinase inhibitor APRin from Pseudomonas aeruginosa

2008 ◽  
Vol 40 (3) ◽  
pp. 213-217 ◽  
Author(s):  
Sengodagounder Arumugam ◽  
Robert D. Gray ◽  
Andrew N. Lane
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Proteinase Inhibitor ◽  
Nmr Structure ◽  
Alkaline Proteinase

scholarly journals Molecular basis for defective secretion of the Z variant of human alpha-1-proteinase inhibitor: secretion of variants having altered potential for salt bridge formation between amino acids 290 and 342.

1989 ◽  
Vol 9 (4) ◽  
pp. 1406-1414 ◽  
Author(s):  
A A McCracken ◽  
K B Kruse ◽  
J L Brown
Keyword(s):  
Amino Acid ◽  
Proteinase Inhibitor ◽  
Conformational Stability ◽  
Point Mutations ◽  
Salt Bridge ◽  
Site Directed Mutagenesis ◽  
Wild Type ◽  
Functional Importance ◽  
Bridge Formation ◽  
Cos Cells

Human alpha-1-proteinase inhibitor (A1PI) deficiency, associated with the Z-variant A1PI (A1PI/Z) gene, results from defective secretion of the inhibitor from the liver. The A1PI/Z gene exhibits two point mutations which specify amino acid substitutions, Val-213 to Ala and Glu-342 to Lys. The functional importance of these substitutions in A1PI deficiency was investigated by studying the secretion of A1PI synthesized in COS cells transfected with A1PI genes altered by site-directed mutagenesis. This model system correctly duplicates the secretion defect seen in individuals homozygous for the A1PI/Z allele and shows that the substitution of Lys for Glu-342 alone causes defective secretion of A1PI. The substitution of Lys for Glu-342 eliminates the possibility for a salt bridge between residues 342 and 290, which may decrease the conformational stability of the molecule and thus account for the secretion defect. However, when we removed the potential to form a salt bridge from the wild-type inhibitor by changing Lys-290 to Glu (A1PI/SB-290Glu), secretion was not reduced to the 19% of normal level seen for A1PI/Z-342Lys; in fact, 75% of normal secretion was observed. When the potential for salt bridge formation was returned to A1PI/Z-342Lys by changing Lys-290 to Glu, only 46% of normal secretion was seen. These data indicate that the amino acid substitution at position 342, rather than the potential to form the 290-342 salt bridge, is the critical alteration leading to the defect in A1PI secretion.

scholarly journals Alkaline Proteinase Inhibitor ofPseudomonas aeruginosa

2003 ◽  
Vol 278 (28) ◽  
pp. 25952-25957 ◽  
Author(s):  
Rhona E. Feltzer ◽  
John O. Trent ◽  
Robert D. Gray
Keyword(s):  
Proteinase Inhibitor ◽  
Alkaline Proteinase

Amino Acid Sequence of an Alkaline Proteinase Inhibitor (Streptomyces Subtilisin Inhibitor) from Streptomyces albogriseolus S-32531

1974 ◽  
Vol 76 (6) ◽  
pp. 1191-1209 ◽  
Author(s):  
Tokuji IKENAKA ◽  
Shoji ODANI ◽  
Mineko SAKAI ◽  
Yoko NABESHIMA ◽  
Sakae SATO ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Proteinase Inhibitor ◽  
Alkaline Proteinase ◽  
Subtilisin Inhibitor
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