DOI: 10.1002/pro.5560040203. Rearranging the domains of pepsinogen

10.1002/pro.5560040203

Crossref journal-article

Wiley

Protein Science (311)

Abstract

AbstractMost eukaryotic aspartic protease zymogens are synthesized as a single polypeptide chain that contains two distinct homologous lobes and a pro peptide, which is removed upon activation. In pepsinogen, the pro peptide precedes the N‐terminal lobe (designated pep) and the C‐terminal lobe (designated sin). Based on the three‐dimensional structure of pepsinogen, we have designed a pepsinogen polypeptide with the internal rearrangement of domains from pro‐pep‐sin (native pepsinogen) to sin‐pro‐pep. The domain‐rearranged zymogen also contains a 10‐residue linker designed to connect sin and pro domains. Recombinant sin‐pro‐pep was synthesized in Escherichia coli, refolded from 8 M urea, and purified. Upon acidification, sin‐pro‐pep autoactivates to a two‐chain enzyme. However, the emergence of activity is much slower than the conversion of the single‐chain zymogen to a two‐chain intermediate. In the activation of native pepsinogen and sin‐pro‐pep, the pro region is cleaved at two sites between residues 16P and 17P and 44P and 1 successively, and complete activation of sin‐pro‐pep requires an additional cleavage at a third site between residues IP and 2P. In pepsinogen activation, the cleavage of the first site is rate limiting because the second site is cleaved more rapidly to generate activity. In the activation of sin‐pro‐pep, however, the second site is cleaved slower than the first, and cleavage of the third site is the rate limiting step. The reason for these differences is the result of the presence of activation intermediates bearing pro peptide 1P‐16P, which is still covalently attached to the sin domain after the first and second cleavages. This peptide is known to have affinity to the enzyme moiety. Its presence apparently prevents the full expression of proteolytic activity, which catalyzes the cleavage of sites 2 and 3. A mechanism of intramolecular cleavage of site 1 is proposed that involves the local conformational change only near site 1 in the N‐terminal region of the pro peptide.

Bibliography

Lin, X., Koelsch, G., Loy, J. A., & Tang, J. (1995). Rearranging the domains of pepsinogen. Protein Science, 4(2), 159â166. Portico.

Authors 4

Xinli Lin (first)
Gerald Koelsch (additional)
Jeffrey A. Loy (additional)
Jordan Tang (additional)

References 18 Referenced 15

10.1016/S0021-9258(19)45033-3
10.1146/annurev.bb.19.060190.001201
10.1042/bj2090355
10.1042/bj1530141 / Biochem J / Conversion of pepsinogen into pepsin is not a one‐step process by Dykes CW (1976)
10.1016/S0021-9258(19)40044-6 / J Biol Chem / Enzymic properties of thermopsin by Fusek M (1990)
10.1002/prot.340130102
10.1093/oxfordjournals.jbchem.a122610 / J Biochem (Tokyo) / Difference of activation processes and structure of activation peptides in human pepsinogen A and progastricsin by Kagayama T (1989)
10.1016/S0021-9258(18)41920-5 / J Biol Chem / Enzymic activities of two‐chain pepsinogen, two‐chain pepsin, and the aminoterminal lobe of pepsinogen by Lin X (1992)
10.1002/pro.5560020903
10.1016/S0021-9258(18)83769-3
10.1016/0003-2697(76)90166-4
10.1126/science.158.3809.1638
10.1111/j.1432-1033.1993.tb18228.x
10.1016/S0021-9258(19)41649-9 / J Biol Chem / Conversion of pepsinogen to pepsin by Sanny CG (1975)
10.1016/0076-6879(90)85008-C
10.1038/271618a0
10.1002/jcb.240330106
10.1021/bi00453a036

Dates

Type	When
Created	15 years, 2 months ago (June 29, 2010, 4:44 a.m.)
Deposited	1 year, 10 months ago (Oct. 26, 2023, 5:30 p.m.)
Indexed	2 months ago (July 4, 2025, 6:07 a.m.)
Issued	30 years, 7 months ago (Feb. 1, 1995)
Published	30 years, 7 months ago (Feb. 1, 1995)
Published Online	16 years, 8 months ago (Dec. 31, 2008)
Published Print	30 years, 7 months ago (Feb. 1, 1995)

Funders 0

None

BibTeX

@article{Lin_1995, title={Rearranging the domains of pepsinogen}, volume={4}, ISSN={1469-896X}, url={http://dx.doi.org/10.1002/pro.5560040203}, DOI={10.1002/pro.5560040203}, number={2}, journal={Protein Science}, publisher={Wiley}, author={Lin, Xinli and Koelsch, Gerald and Loy, Jeffrey A. and Tang, Jordan}, year={1995}, month=feb, pages={159–166} }

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