Proteolytic activity of pseudotrypsin

Keil-Dlouhá, V.; Zylber, N.; Imhoff, Jean-Marie; Tong, Nguyen Thanh; Keil, B.

doi:10.1016/0014-5793(71)80373-3

Cited by 113 publications

(88 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Efforts to prevent this inherent chymotryptic-like activity of trypsin have failed in agreement with the experiences of other investigators [43,45]. We presume that this non-tryptic specificity may reflect the presence of $-trypsin [46]. The exchange protein contains four tryptophan residues of which three residues are assigned to the N-terminal half (position 30, 81 and 101).…”

Section: Discussionsupporting

confidence: 83%

The Primary Structure of the Phosphatidylcholine‐Exchange Protein from Bovine Liver

Moonen

Akeroyd

Westerman

et al. 1980

European Journal of Biochemistry

View full text Add to dashboard Cite

The phosphatidylcholine exchange protein from bovine liver consists of a single polypeptide chain and has a blocked N terminus. The protein contains an estimated 244 amino acid residues in accordance with a determined molecular weight of 28 000. The protease from mouse submaxillaris gland cleaved the citraconylated and S-carboxymethylated derivative of the exchange protein at one specific site (Argl4-GluI5) close to the N terminus. Analysis of the two resulting peptides showed that N-acetyl-methionine was the N-terminal residue and gave the sequence of the first 41 residues.The modified protein was also fragmented with the protease from Staphylococcus aureus. The peptides isolated represented 88 of the protein; their sequences were determined by manual and automated Edman degradation. Alignment of a number of these peptides gave the complete sequence of the N-terminal half up to position 322.Phospholipid exchange proteins catalyze the transfer of phospholipids between membranes. A number of these proteins, different in molecular weight, isoelectric point and transfer specificity, have been isolated from bovine liver [l], heart [2,3] and brain [4,5], rat liver [6 -81, small intestinal mucosa [9] and hepatoma [lo], sheep lung [ l l ] and potato tuber [12]. To date, three general classes of exchange proteins can be distinguished, that is the phosphatidylcholinespecific exchange protein [1,6,9,11], the exchange proteins that display a great preference for phosphatidylinositol [4 -6,9] and the non-specific exchange proteins [8, lo]. These proteins range in molecular weight between 10000 and 35000 and in isoelectric point between pH 5 and 9.

show abstract

Section: Discussionsupporting

confidence: 83%

The Primary Structure of the Phosphatidylcholine‐Exchange Protein from Bovine Liver

Moonen

Akeroyd

Westerman

et al. 1980

European Journal of Biochemistry

View full text Add to dashboard Cite

show abstract

“…However, this did not result in a less complex pattern of spots in the peptide maps, nor did the use of different commercial Tos-Phe-CH,Cl-treated trypsins for digestion. A prolonged treatment of cytochrome L' with commercial Tos-Phe-CH,Cl-treated trypsin with the intention of quantitatively cleaving it at all its lysine residues gave an even more complex pattern of spots due to atypical cleavage by autolytically created $-trypsin [53]. Therefore throughout the investigations the standardized conditions of tryptic digestions described were used.…”

Section: Resultsmentioning

confidence: 99%

Change of Cytochrome c Structure during Development of the Mouse

Hennig

1975

European Journal of Biochemistry

View full text Add to dashboard Cite

The structure of cytochrome c during mouse development is investigated. For this purpose the amino acid sequence of cytochrome c of the adult mouse had to be determined. The structure of cytochrome c of adult differentiated mouse cells differs in two amino acid residues from the known amino acid sequence of rabbit cytochrome c. No indication of different forms of cytochrome c in the adult differentiated cells was obtained. The structure of cytochrome c from 11.5‐day‐old mouse embryos is identical with that of adult mouse tissues. Since germ cells after meiotic division are the immediate precursors of a new individual, the structure of cytochrome c from sperm‐containing mice testes was investigated. By means of chromatography of the cytochrome c and of peptide maps and amino acid analyses of its tryptic peptides, it is shown that mouse testis contains two isocytochromes c in about equal amount. The structure of one of these two isocytochromes c is identical with the structure of the adult‐type cytochrome c of mouse. The testis‐specific cytochrome c, which is assumed to be located in the sperm cells, differs in 13 of its 104 amino acid residues from the adult‐type cytochrome c. From comparison of the primary and the spatial structures of the adult‐type and the sperm‐type isocytochromes c with the known structures of cytochrome c of more than 65 different species it is concluded that the duplication of the cytochrome c structural gene, causing the existence of the two ontogenetic‐specific isocytochromes c in mouse, has occurred early in the evolution of eucaryotes.

show abstract

“…In our previous work we were able to show that this particular cleavage destroys the "chymotryptic" active site of the trypsin molecule [2]. We have also proved that only one electrostatic bond exists between Asp-177 of trypsin and Lys-15 of the inhibitor and that the other interactions are due to hydrophobic forces [1 ]. Alkylation of His-46 in a-and/3-trypsin with TLCK leads to the same chemical modification resulting in the inactivation of the specific tryptic active site which is responsible for cleavage of positively charged substrates.…”

Section: Resultsmentioning

confidence: 67%

“…This is an interaction of hydrophobic character. Consequently, during the complex formation between trypsin and pancreatic inhibitor the inhibition of two active sites occurs: 1) the inhibition of the specific active site of trypsin by an electrostatic interaction between Asp-177 of trypsin and Lys-15 of the inhibitor [ 1], 2) the inhibition of the nonspecific "chymotryptic" active site of trypsin through a hydrophobic interaction between hydrophobic residues of trypsin and the inhibitor. …”

Section: Resultsmentioning

confidence: 99%

“…The study of the complex between a-, t-and ~k-trypsin and the pancreatic inhibitor showed that only one electrostatic bond is involved in complex formation between trypsin and BPTI* [1] -the bond between Asp-177, which is a specificity site carboxyl group of trypsin, binding positively charged substrates, and Lys-15 of pancreatic inhibitor. We were able to show that hydrophobic forces play an important role in the interaction between trypsin and BPTI.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

On the complex formation between basic pancreatic trypsin inhibitor and TLCK‐, TPCK‐derivatives of β‐trypsin

Imhoff

Keil-Dlouhá

1971

FEBS Letters

View full text Add to dashboard Cite

The study of the interaction of the pancreatic inhibitor with different alkylated derivatives of a-and E-trypsin shows that: 1) TLCK~-trypsin forms a complex with pancreatic inhibitor in tris buffer and tris-ethano140% system. 2) TLCK-a-trypsin and TLCK-TPCK-fl-trypsin have lost their ability to complex formation with pancreatic inhibitor. TLCK-ot-trypsin and TLCK-TPCK-#-trypsin are derivatives in which the "chymotryptic" active site is destroyed. The results presented in this paper prove the participation of the "chymotrypic'" active site in the interaction between trypsin and pancreatic inhibitor. This is the second interaction beside that of the electrostatic bond between Asp-117 of trypsin and Lys-15 of the inhibitor which we proved earlier.

show abstract

Proteolytic activity of pseudotrypsin

Cited by 113 publications

References 10 publications

The Primary Structure of the Phosphatidylcholine‐Exchange Protein from Bovine Liver

The Primary Structure of the Phosphatidylcholine‐Exchange Protein from Bovine Liver

Change of Cytochrome c Structure during Development of the Mouse

On the complex formation between basic pancreatic trypsin inhibitor and TLCK‐, TPCK‐derivatives of β‐trypsin

Contact Info

Product

Resources

About