Structure of the complex formed by bovine trypsin and bovine pancreatic trypsin inhibitor

Huber, Robert; Kukla, D.; Bode, Wolfram; Schwager, P.; Bartels, Klaus; Deisenhofer, Johann; Steigemann, W.

doi:10.1016/0022-2836(74)90163-6

Cited by 560 publications

(226 citation statements)

References 21 publications

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“…Secondly, the shift in charge state is also very significant as Table 1 K~ values of trypsin-inhibitor complexes Inhibitor BPTI [10] RcamBPTI [9] SBTI [17,18] K15V BPTI [14] Ki (M) 6.0 X 10 -14 1.3 × 10 -9 1 × 10 -11 does not inhibit .~ A.E. Kraunsoe et al/FEBS Letters 396 (1996) tein proteinase inhibitor complexes suggest that 10-15 residues of the inhibitor form close contacts with 17 29 residues of the proteinase.…”

Section: Jae Kraunsoe Et Al/febs Letters 396 (1996) 108-112mentioning

confidence: 99%

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An investigation of the binding of protein proteinase inhibitors to trypsin by electrospray ionization mass spectrometry

Kraunsoe

Aplin

Green³

et al. 1996

FEBS Letters

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The binding of BPTI and SBTI with trypsin has been investigated by ESI MS, using the mutant K15V-BPTI and the chemically modified RcamBPTI as controls. Although high cone voltages (+80 V) produce sharp spectra of BPTI, RcamBPTI, SBTI and trypsin alone, the complexes of BPTI, RcamBPTI and SBTI with trypsin undergo partial dissociation due to collisional activation. At lower cone voltages (+40 V) these non-covalent complexes are stable. The charge distribution on the trypsin and the inhibitors produced by gas phase dissociation of the complexes are markedly different from those of the components alone, indicating that ESI MS provides a novel probe for exploring the ionic interactions at the contact surface of proteins. Moreover, by determining the cone voltage at which the gas phase dissociation of complexes occurs it may be possible to use ESI MS to compare the binding energies of closely related complexes.

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Section: Jae Kraunsoe Et Al/febs Letters 396 (1996) 108-112mentioning

confidence: 99%

“…The crystal structures of the BPTI complexes with 13-trypsin and porcine kallikrein A have been solved [8,9]. These have shown that the region of interaction, as for many proteinase inhibitors, is between the active site cleft of the proteinase and an external loop region of the BPTI.…”

Section: Introductionmentioning

confidence: 99%

An investigation of the binding of protein proteinase inhibitors to trypsin by electrospray ionization mass spectrometry

Kraunsoe

Aplin

Green³

et al. 1996

FEBS Letters

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“…As the principal template for construction of both forms of HBP the structure of human neutrophil elastase (PDB code 1HNE; Navia et al, 1989) complexes of BPTI with trypsin (PDB code 2PTC; Huber et al, 1974) and kallikrein (PDB code 2KAI; Chen and Bode, 1983) were used in guiding the construction of the model for the complex of hHBP with BPTI. The only regions in the two HBPs where the model construction required special attention are the section that includes residues 214-226, and in particular, near residue 220.…”

Section: Modeling Of Hhbp and Phbpmentioning

confidence: 99%

“…Its primary amino acid sequence (Kassel et al, 1965) and genomic sequence (Anderson and Kingston, 1983), as well as its three-dimensional structure (Huber et al, 1974), have been determined. BPTI and closely related proteinase inhibitory proteins have been found in high amounts in organs from ruminants where they have been located in the mast cells (Fritz et al, 1979).…”

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confidence: 99%

Binding of bovine pancreatic trypsin inhibitor to heparin binding protein/CAP37/azurocidin

Petersen

Birktoft

Flodgaard

1993

European Journal of Biochemistry

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Heparin-binding protein (HBP; also known as CAP37 or azurocidin) is a member of the serine proteinase family. Evolution, however, has reverted this protein into a non-proteolytic form by mutation of two of the three residues of the active-site triad. Although proteolytically inactive, the human heparin-binding protein (hHBP) is still capable of binding bovine pancreatic trypsin inhibitor (BPTI). This was demonstrated by affinity chromatography to BPTI immobilized on a solid matrix and by studies on plasmin inhibition kinetics. hHBP competes with plasmin for BPTI and this effect on plasmin inhibition has been analyzed in terms of a kinetic model. A dissociation constant, Kd = 0.1 pM, was found for the interaction between BPTI and hHBP. The hHBP provides an example of a serine proteinase which has lost its catalytic function by reverting residues of the active center while still preserving its capability of specific interactions with Kunitz inhibitors. pHBP, the porcine counterpart to hHBP, on the other hand, was incapable of BPTI binding. The structural basis for the BPTI binding to the human protein and the species difference is discussed in terms of putative three-dimensional structures of the proteins derived by comparative molecular modelling methods.

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“…5, one region in the middle of the inhibitor molecule differs to the same extent or more between the two cobra species than between them and the pancreatic inhibitor. However, in relation to the known tertiary structure of the pancreatic inhibitor [8], it may be noted that the contact site with serine proteases is largely conserved. Of positions suggested to interact with the enzyme, only the one at P3' and to some extent the one at P2' are markedly different in the Nuju nuju nuju inhibitor (Table I), which can be functionally interpreted in terms of strong inhibition (below).…”

Section: Resultsmentioning

confidence: 99%

Primary structure and functional properties of cobra (Naja naja naja) venom Kunitz‐type trypsin inhibitor

Shafqat

Beg

Yin

et al. 1990

European Journal of Biochemistry

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A trypsin inhibitor from the venom of the cobra Nuja nuju nuja has been isolated by a single step of reversephase high-performance liquid chromatography. The protein strongly inhibits trypsin (Ki = 3.5 pM). The primary structure was determined by peptide analysis of the [ ''C]carboxymethylated inhibitor. The 57-residue polypeptide chain belongs to the family of Kunitz-type inhibitors, and exhibits 42% residue identity with bovine pancreatic trypsin inhibitor. The structure shows only 70% identity with the corresponding peptide from the Capa cobra (Naju neviu), establishing that the inhibitor molecule exhibits extensive variations. Functionally, a basic residue at position P3' correlates with strong inhibition.Basic protease inhibitors of the serine proteases are abundant in nature and appear to be present in all forms of life [l]. Different types have been isolated and extensively studied. Based on different properties, they have been classified into at least seven families [2].Studies of snake venoms reveal the presence of several protease inhibitors, particularly from Elapidae and Viperidae snakes. These inhibitors belong to the protein family of pancreatic trypsin inhibitors of the Kunitz type [3]. The amino acid sequences of the venom inhibitors from Viperu russelli (Russell's viper) [4], Nuju neviu (Cape cobra) [5], Dendroaspis polylepis (black mamba) [6], Vipera ummodytes (long-nosed viper) [7] and Hemachatus haemuchutus (Ringhalls cobra) [5] have been characterized. These analyses, correlated with the known tertiary structure of the pancreatic inhibitor [S], have facilitated understanding of the structure/function relationships for the inhibitors. Comparisons show conservation of residues at both the site of the main contacts with serine proteases [9] and the region of weak contacts [S]. The snake venom inhibitors can inhibit the activities of serine proteases, including bovine trypsin, kallikrein and plasmin [7].During investigation of the venom of the nominate race of cobra, Naja naja naja (from Pakistan), we have isolated a highly potent trypsin inhibitor of this venom. In this work, we report a single-step purification of the inhibitor on reversephase HPLC, enzymatic parameters for the protease inhibition, and the primary structure of the inhibitor, explaining its potency and antitrypsin activity.

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Structure of the complex formed by bovine trypsin and bovine pancreatic trypsin inhibitor

Cited by 560 publications

References 21 publications

An investigation of the binding of protein proteinase inhibitors to trypsin by electrospray ionization mass spectrometry

An investigation of the binding of protein proteinase inhibitors to trypsin by electrospray ionization mass spectrometry

Binding of bovine pancreatic trypsin inhibitor to heparin binding protein/CAP37/azurocidin

Primary structure and functional properties of cobra (Naja naja naja) venom Kunitz‐type trypsin inhibitor

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