The role of arginines in stabilizing the active open‐lid conformation of Rhizomucor miehei lipase
Abstract Molecular dynamics simulations for the lid covering the active site of Rhizomucor miehei lipase [EC 3.1.1.3] postulated that, among other interactions, Arg86 in the lid stabilized the open‐lid conformation of the protein by multiple hydrogen bonding to the protein surface. Chemical modifica...
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crwiley:10.1007/bf02535993 2024-06-23T07:47:44+00:00 The role of arginines in stabilizing the active open‐lid conformation of Rhizomucor miehei lipase Holmquist, Mats Norin, Martin Hult, Karl 1993 http://dx.doi.org/10.1007/bf02535993 https://onlinelibrary.wiley.com/doi/full/10.1007/BF02535993 en eng Wiley http://onlinelibrary.wiley.com/termsAndConditions#vor Lipids volume 28, issue 8, page 721-726 ISSN 0024-4201 1558-9307 journal-article 1993 crwiley https://doi.org/10.1007/bf02535993 2024-06-11T04:45:36Z Abstract Molecular dynamics simulations for the lid covering the active site of Rhizomucor miehei lipase [EC 3.1.1.3] postulated that, among other interactions, Arg86 in the lid stabilized the open‐lid conformation of the protein by multiple hydrogen bonding to the protein surface. Chemical modification of arginine residues in R. miehei lipase with 1,2‐cyclohexanedione or phenylglyoxal resulted in residual activities in the hydrolysis of tributyrin of 66 and 46%, respectively. Tryptic maps of native and phenylglyoxal‐reacted R. miehei lipase showed that Arg86 was the residue modified most, when the lipase was inhibited to the greatest extent. Guanidine, a structural analog to an arginine side chain, inhibited both the natibe enzyme and the arginine‐modified enzymes, resulting in residual activities of 26% as compared to the native enzyme. The inhibition was not an effect of enzyme denaturation. The native enzyme was also inhibited by 1‐ethylguanidine, benzamidine and urea, but to a lesser degree than by guanidine. Lipases from Humicola lanuginosa and porcine pancreas in 100 mM guanidine showed residual activities of 88 and 70%, respectively. The lipases from Candida antarctica, C. rugosa, Pseudomonas cepacia and P. fluoresences were not inhibited by guanidine. The inhibition of R. miehei lipase by structural analogs of the arginine side chain and after chemical modification of arginine residues suggest a role of an arginine residue in stabilizing the active open‐lid conformation of the enzyme. Article in Journal/Newspaper Antarc* Antarctica Wiley Online Library Rugosa ENVELOPE(-61.250,-61.250,-62.633,-62.633) Lipids 28 8 721 726 |
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English |
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Abstract Molecular dynamics simulations for the lid covering the active site of Rhizomucor miehei lipase [EC 3.1.1.3] postulated that, among other interactions, Arg86 in the lid stabilized the open‐lid conformation of the protein by multiple hydrogen bonding to the protein surface. Chemical modification of arginine residues in R. miehei lipase with 1,2‐cyclohexanedione or phenylglyoxal resulted in residual activities in the hydrolysis of tributyrin of 66 and 46%, respectively. Tryptic maps of native and phenylglyoxal‐reacted R. miehei lipase showed that Arg86 was the residue modified most, when the lipase was inhibited to the greatest extent. Guanidine, a structural analog to an arginine side chain, inhibited both the natibe enzyme and the arginine‐modified enzymes, resulting in residual activities of 26% as compared to the native enzyme. The inhibition was not an effect of enzyme denaturation. The native enzyme was also inhibited by 1‐ethylguanidine, benzamidine and urea, but to a lesser degree than by guanidine. Lipases from Humicola lanuginosa and porcine pancreas in 100 mM guanidine showed residual activities of 88 and 70%, respectively. The lipases from Candida antarctica, C. rugosa, Pseudomonas cepacia and P. fluoresences were not inhibited by guanidine. The inhibition of R. miehei lipase by structural analogs of the arginine side chain and after chemical modification of arginine residues suggest a role of an arginine residue in stabilizing the active open‐lid conformation of the enzyme. |
format |
Article in Journal/Newspaper |
author |
Holmquist, Mats Norin, Martin Hult, Karl |
spellingShingle |
Holmquist, Mats Norin, Martin Hult, Karl The role of arginines in stabilizing the active open‐lid conformation of Rhizomucor miehei lipase |
author_facet |
Holmquist, Mats Norin, Martin Hult, Karl |
author_sort |
Holmquist, Mats |
title |
The role of arginines in stabilizing the active open‐lid conformation of Rhizomucor miehei lipase |
title_short |
The role of arginines in stabilizing the active open‐lid conformation of Rhizomucor miehei lipase |
title_full |
The role of arginines in stabilizing the active open‐lid conformation of Rhizomucor miehei lipase |
title_fullStr |
The role of arginines in stabilizing the active open‐lid conformation of Rhizomucor miehei lipase |
title_full_unstemmed |
The role of arginines in stabilizing the active open‐lid conformation of Rhizomucor miehei lipase |
title_sort |
role of arginines in stabilizing the active open‐lid conformation of rhizomucor miehei lipase |
publisher |
Wiley |
publishDate |
1993 |
url |
http://dx.doi.org/10.1007/bf02535993 https://onlinelibrary.wiley.com/doi/full/10.1007/BF02535993 |
long_lat |
ENVELOPE(-61.250,-61.250,-62.633,-62.633) |
geographic |
Rugosa |
geographic_facet |
Rugosa |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
Lipids volume 28, issue 8, page 721-726 ISSN 0024-4201 1558-9307 |
op_rights |
http://onlinelibrary.wiley.com/termsAndConditions#vor |
op_doi |
https://doi.org/10.1007/bf02535993 |
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Lipids |
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28 |
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8 |
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721 |
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726 |
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1802651894982639616 |