Enhanced Plasmin Inhibition by a Reactive Center Lysine Mutant of the Kunitz-type Protease Inhibitor Domain of the Amyloid β-Protein Precursor

Nostrand, William E. Van; Siegel, Robert S.; Wagner, Steven L.; Raschke, William C.

doi:10.1074/jbc.270.39.22827

Cited by 24 publications

(25 citation statements)

References 23 publications

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“…In reported specificity studies of target plasma proteases inhibited by PN2/AbPP and by PN2KPI, there is a general consensus that neither thrombin (21,54) nor FXIIa (21,40) is inhibited by PN2KPI, in agreement with the present studies which show no inhibition by thrombin and a K i $ 7610 nM for FXIIa (Table I). There is also a consensus that both FXIa (18,21,40,55) and trypsin (18,21,40) are potently inhibited by PN2KPI with K i values of 0.252.7 nM and 0.020.42 nM, respectively, i.e.…”

Section: Fxia and Trypsin Inhibition By Pn2kpi And Bptisupporting

confidence: 81%

“…Human Kunitz-type inhibitors are also known to inhibit nonhuman proteinases. For example, bovine trypsin and bovine chymotrypsin are inhibited by human TFPI (38), TFPI 2 (39) and PN2KPI (21,40), whereas mouse NGF-gamma is inhibited by human PN2KPI (21). In addition to these biochemical studies, three-dimensional structures of cross-species enzyme/inhibitor complexes have also been documented, including the crystal structures of human PN2KPI in complex with bovine trypsin, bovine chymotrypsin (41), and rat anionic trypsin (42) as well as BPTI in complex with human mesotrypsin and human cationic trypsin (33).…”

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confidence: 99%

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Mechanisms and specificity of factor XIa and trypsin inhibition by protease nexin 2 and basic pancreatic trypsin inhibitor

Navaneetham

Sinha²,

Walsh

2010

The Journal of Biochemistry

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Factor XIa (FXIa) inhibition by protease nexin-2 (PN2KPI) was compared with trypsin inhibition by basic pancreatic trypsin inhibitor (BPTI). PN2KPI was a potent inhibitor of FXIa (K i $ 0.81 nM) and trypsin (K i $ 0.03 nM), but not of other coagulation proteases (thrombin, FVIIa, FIXa, FXa, FXIIa, plasmin, kallikrein, K i 4185 nM). PN2KPI was $775-fold more potent than BPTI in FXIa inhibition, but both exhibited similar potencies against trypsin. Studies of FXIa and trypsin inhibition by PN2KPI and BPTI and P1 site swap mutants (PN2KPI-R15 K, BPTI-K15 R) demonstrated that FXIa inhibition by PN2KPI and P1 site swap mutants and trypsin inhibition by PN2KPI and BPTI conform to a single-step, slow equilibration inhibitory mechanism, whereas FXIa-inhibition by BPTI follows a classical, competitive inhibitory mechanism. Mutation of P1 impaired FXIa inhibition by PN2KPI-R15 K $14-fold, enhanced FXIa inhibition by BPTI-K15 R $150-fold, and had no effect on trypsin inhibition. Arginine at the P1 site of either PN2KPI or BPTI confers high affinity and specificity for FXIa, whereas either arginine or lysine suffices for trypsin inhibition. Thus, PN2KPI is a highly specific inhibitor of FXIa among coagulation enzymes, but the flexibility of trypsin renders it susceptible to inhibition by both wild-type and mutant forms of PN2KPI and BPTI.Keywords: BPTI/FXIa/inhibition mechanism/ protease nexin 2 Kunitz domain/trypsin.Abbreviations: BMMY, buffered medium containing methanol and yeast nitrogen base; BMGY, buffered medium containing glycerol and yeast nitrogen base; BPTI, basic (or bovine) pancreatic trypsin inhibitor; PN2KPI, protease nexin 2 Kunitz protease inhibitory domain; PN2, protease nexin 2; KPI, Kunitz protease

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Section: Fxia and Trypsin Inhibition By Pn2kpi And Bptisupporting

confidence: 81%

mentioning

confidence: 99%

Mechanisms and specificity of factor XIa and trypsin inhibition by protease nexin 2 and basic pancreatic trypsin inhibitor

Navaneetham

Sinha²,

Walsh

2010

The Journal of Biochemistry

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“…1), the arginine residue in position P1 in C5 and APPI could potentially give them an antiprotease activity. This prediction was confirmed for APPI (Van Nostrand et al, 1995) but not for C5. C5 has no inhibitory activity against trypsin, chymotrypsin, thrombin, kallikrein, neutrophile elastase, cathepsin G, factor VIIa or Xa (Petersen, unpublished results) or against mast cell chymase (Mayer et al, 1994).…”

Section: Introductionsupporting

confidence: 69%

1.2 Å Refinement of the Kunitz-Type Domain from the α3 Chain of Human Type VI Collagen

Merigeau

Arnoux

Pérahia

et al. 1998

Acta Crystallogr D Biol Cryst

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The recombinant Kunitz-type domain (C5) of human collagen cr3(VI) chain was previously described at 1.6 ,A resolution at room temperature. By changing the crystallization conditions and using synchrotron radiation, we are able to record diffraction data to 1.2,~ resolution for crystals of the same space group at 291 K. The protein-water-ion model has been refined anisotropically against these new data using the program SHELXL93; the results converged to an R factor of 15.0%, with all data between 7 and 1.2,4,. The final electron-density map reveals a clear chain tracing with a few disordered residues and five residues out of 58 that present alternate conformations. The Cys14-Cys38 bond presents the less frequently observed left-hand conformation (Xl = -60°) • The solvent molecules and a phosphate ion are well ordered with an average B of 38 ~2. The high-resolution structure reveals the N and C termini which were missing from the 1.6 A structure.

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“…The binding of APP leads to the presence of the Kunitz-type protease inhibitor (KPI) domain, which is part of most of the APP isoforms. The KPI domain inhibits coagulant factors XIa and IXa (Van Nostrand et al, 1995), and Aβ fibrils enhance the anticoagulant property of APP (Wagner et al, 2000). As a consequence, an anticoagulant environment is created, leading to an increased chance of hemorrhages.…”

Section: Aβ Fibril Assembly At Cell Surfacesmentioning

confidence: 99%

Amyloid β in hereditary cerebral hemorrhage with amyloidosis-Dutch type

Kamp

Moursel²,

Haan³

et al. 2014

Reviews in the Neurosciences

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Enhanced Plasmin Inhibition by a Reactive Center Lysine Mutant of the Kunitz-type Protease Inhibitor Domain of the Amyloid β-Protein Precursor

Cited by 24 publications

References 23 publications

Mechanisms and specificity of factor XIa and trypsin inhibition by protease nexin 2 and basic pancreatic trypsin inhibitor

Mechanisms and specificity of factor XIa and trypsin inhibition by protease nexin 2 and basic pancreatic trypsin inhibitor

1.2 Å Refinement of the Kunitz-Type Domain from the α3 Chain of Human Type VI Collagen

Amyloid β in hereditary cerebral hemorrhage with amyloidosis-Dutch type

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