Solid-state nuclear magnetic resonance investigation of solvent dependence of tyrosyl ring motion in an enzyme
Tyrosyl ring motions in alpha-lytic protease were investigated by solid-state deuterium nuclear magnetic resonance (NMR) spectroscopy in lyophilized enzyme powder, in powder suspended in organic solvents, and in aqueous crystals. Ring flipping rates were determined by examining deuterium quadrupole...
Saved in:
Published in | Biotechnology and bioengineering Vol. 42; no. 1; p. 87 |
---|---|
Main Authors | , , |
Format | Journal Article |
Language | English |
Published |
United States
05.06.1993
|
Online Access | Get more information |
Cover
Loading…
Abstract | Tyrosyl ring motions in alpha-lytic protease were investigated by solid-state deuterium nuclear magnetic resonance (NMR) spectroscopy in lyophilized enzyme powder, in powder suspended in organic solvents, and in aqueous crystals. Ring flipping rates were determined by examining deuterium quadrupole echo line shapes. Of the four Tyr residues in the enzyme, one was flipping at the slow (< or =10(3) s(-1)) and one at the fast (> or =10(7) s(-1)) exchange limit of the line shape experiment in all the environments tested. Flipping rates of the remaining two Tyr residues depended markedly on the solvent, with the lowest flipping rates (< or =10(3) s(-1) for both residues) observed in the enzyme powder, whether dry or suspended in hydrophobic tert-butyl methyl ether. In hydrophilic dioxane and acetonitrile, the mobility of these residues increased to 10(4) and 10(5) s(-1). The latter rate rose further to 10(6) s(-1) in the hydrated hydrophilic solvents and to > or =10(7) s(-1) in aqueous crystals. The deuterium spectrum of native alpha-lytic protease was compared with that of the enzyme whose active center was covalently modified with an inhibitor, which binds next to Tyr-123, constraining its ring. This experiment revealed that water addition to acetonitrile specifically increased the flipping rate of this active center residue. Librational motions ("wobbling"), estimated by their effect on spin-lattice relaxation times, were slowest in the anhydrous solvents, intermediate in the hydrated solvents, and fastest in the aqueous crystals. Thus, alpha-lytic protease is more rigid in organic solvents than in water, as judged by mobility of its tyrosyl residues. Water stripping by hydrophilic solvents did not increase enzyme rigidity, nor were there clear correlations between mobility and either enzymatic activity or solvent dielectric constant. |
---|---|
AbstractList | Tyrosyl ring motions in alpha-lytic protease were investigated by solid-state deuterium nuclear magnetic resonance (NMR) spectroscopy in lyophilized enzyme powder, in powder suspended in organic solvents, and in aqueous crystals. Ring flipping rates were determined by examining deuterium quadrupole echo line shapes. Of the four Tyr residues in the enzyme, one was flipping at the slow (< or =10(3) s(-1)) and one at the fast (> or =10(7) s(-1)) exchange limit of the line shape experiment in all the environments tested. Flipping rates of the remaining two Tyr residues depended markedly on the solvent, with the lowest flipping rates (< or =10(3) s(-1) for both residues) observed in the enzyme powder, whether dry or suspended in hydrophobic tert-butyl methyl ether. In hydrophilic dioxane and acetonitrile, the mobility of these residues increased to 10(4) and 10(5) s(-1). The latter rate rose further to 10(6) s(-1) in the hydrated hydrophilic solvents and to > or =10(7) s(-1) in aqueous crystals. The deuterium spectrum of native alpha-lytic protease was compared with that of the enzyme whose active center was covalently modified with an inhibitor, which binds next to Tyr-123, constraining its ring. This experiment revealed that water addition to acetonitrile specifically increased the flipping rate of this active center residue. Librational motions ("wobbling"), estimated by their effect on spin-lattice relaxation times, were slowest in the anhydrous solvents, intermediate in the hydrated solvents, and fastest in the aqueous crystals. Thus, alpha-lytic protease is more rigid in organic solvents than in water, as judged by mobility of its tyrosyl residues. Water stripping by hydrophilic solvents did not increase enzyme rigidity, nor were there clear correlations between mobility and either enzymatic activity or solvent dielectric constant. |
Author | Griffin, R G Klibanov, A M Burke, P A |
Author_xml | – sequence: 1 givenname: P A surname: Burke fullname: Burke, P A organization: Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA – sequence: 2 givenname: R G surname: Griffin fullname: Griffin, R G – sequence: 3 givenname: A M surname: Klibanov fullname: Klibanov, A M |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/18609651$$D View this record in MEDLINE/PubMed |
BookMark | eNo1j0tLAzEUhbOo2Icu3Ur-wNQ8mkxmKUWtUHChrkuSuSmRmZsySQvjr7e-Vofv8HHgzMkEEwIhN5wtOWPizsWyFJqtBONcTMiMMaYrqRoxJfOcP85YG60vyZQbzRqt-Izga-piW-ViC1A8-g7sQHu7RyjR0wFyQoseaMQT5BL3tsSENAWaU3cCLLSFA2AL3865LeOQ8tjRIeKe9ulHjkgtUsDPsYcrchFsl-H6Lxfk_fHhbb2pti9Pz-v7beWVNKIyK94wFhz3wXmnhV3ZptZWBG08d7KxzIRglJK8FlZKDSE0WtTKGWWlcY1YkNvf3cPR9dDuDkPs7TDu_o-LL_dfXFc |
CitedBy_id | crossref_primary_10_1016_S0021_9258_18_31734_4 crossref_primary_10_1002_bit_10536 crossref_primary_10_1016_S1367_5931_99_00055_1 crossref_primary_10_1002_pro_2271 crossref_primary_10_1002_bit_20863 crossref_primary_10_1002_bit_21510 crossref_primary_10_3390_catal8080338 crossref_primary_10_1016_j_jclepro_2016_09_211 crossref_primary_10_1007_s10295_009_0610_3 crossref_primary_10_1021_op300335d crossref_primary_10_1038_374596a0 crossref_primary_10_1016_S0022_2860_01_00770_0 crossref_primary_10_1021_ma991344g crossref_primary_10_1080_10409239891204170 crossref_primary_10_1021_ja806996q crossref_primary_10_1021_js980272o crossref_primary_10_1073_pnas_0601113103 crossref_primary_10_1021_ja970629a crossref_primary_10_1016_S0022_2860_97_00257_3 crossref_primary_10_1016_j_biortech_2012_01_088 crossref_primary_10_1021_cr0404242 crossref_primary_10_3109_10242429709003609 crossref_primary_10_1021_bi960191b crossref_primary_10_1246_bcsj_76_399 crossref_primary_10_1002__SICI_1097_0290_19960220_49_4_399__AID_BIT6_3_0_CO_2_K crossref_primary_10_1038_35051719 crossref_primary_10_1021_jp962116g crossref_primary_10_1039_c0cp00350f crossref_primary_10_3762_bjoc_9_56 crossref_primary_10_1002__SICI_1097_0290_19960105_49_1_87__AID_BIT11_3_0_CO_2_8 crossref_primary_10_1016_j_enzmictec_2018_05_010 crossref_primary_10_1002_chem_201000487 crossref_primary_10_1016_S0141_0229_97_00241_X crossref_primary_10_1016_S0167_4838_98_00086_7 crossref_primary_10_1016_j_biochi_2011_04_017 crossref_primary_10_1073_pnas_0804566105 crossref_primary_10_1016_S0141_0229_00_00240_4 crossref_primary_10_1098_rstb_2004_1505 crossref_primary_10_1007_BF03167173 crossref_primary_10_1002__SICI_1097_0290_19960505_50_3_257__AID_BIT4_3_0_CO_2_F crossref_primary_10_1016_0167_4838_94_90065_5 crossref_primary_10_1002__SICI_1097_0290_19980320_57_6_686__AID_BIT6_3_0_CO_2_H crossref_primary_10_1016_j_bbapap_2004_12_007 crossref_primary_10_1016_S0141_0229_00_00358_6 crossref_primary_10_1021_acs_jpcb_8b06330 crossref_primary_10_1002_1521_3757_20000703_112_13_2312__AID_ANGE2312_3_0_CO_2_4 crossref_primary_10_3109_10242420009015259 crossref_primary_10_1016_j_tetasy_2008_03_021 crossref_primary_10_1021_jp208966k crossref_primary_10_1016_S0141_0229_01_00421_5 |
ContentType | Journal Article |
Copyright | (c) 1993 John Wiley & Sons, Inc. |
Copyright_xml | – notice: (c) 1993 John Wiley & Sons, Inc. |
DBID | NPM |
DOI | 10.1002/bit.260420112 |
DatabaseName | PubMed |
DatabaseTitle | PubMed |
DatabaseTitleList | PubMed |
Database_xml | – sequence: 1 dbid: NPM name: PubMed url: https://proxy.k.utb.cz/login?url=http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed sourceTypes: Index Database |
DeliveryMethod | no_fulltext_linktorsrc |
Discipline | Engineering Chemistry Biology Anatomy & Physiology |
ExternalDocumentID | 18609651 |
Genre | Journal Article |
GroupedDBID | --- -~X .GJ 0R~ 1L6 1OB 1OC 1ZS 23N 33P 3EH 3SF 3WU 4ZD 50Y 50Z 51W 51X 52M 52N 52O 52P 52T 52U 52W 52X 53G 5GY 5RE 5VS 66C 702 7PT 8-0 8-1 8UM AAESR ABCQN ABEML ABJNI ACBWZ ACGFO ACGFS ACIWK ACPRK ACXQS ADKYN ADOZA ADZMN AENEX AEUQT AFBPY AFFNX AFRAH AI. AIAGR ALAGY ALMA_UNASSIGNED_HOLDINGS AMBMR ASPBG ATUGU AVWKF AZFZN BAFTC BDRZF BFHJK BLYAC BNHUX BY8 CS3 D-E D-F DCZOG DR1 DR2 DU5 EBS EJD F04 F5P FEDTE GNP GODZA H.T H.X HBH HHY HHZ HVGLF HZ~ IX1 J0M JPC KQQ LATKE LAW LC2 LC3 LEEKS LH6 LOXES LUTES LW6 MK4 MRFUL N04 N05 N9A NDZJH NF~ NNB NPM O66 O9- OIG P2P P2W P2X PALCI PQQKQ Q.N QRW RBB RIWAO RJQFR RWI RX1 SAMSI SUPJJ TN5 VH1 W8V W99 WBKPD WH7 WIB WIH WJL WNSPC WSB WXSBR XPP XV2 Y6R ZXP ~02 ~KM ~WT |
ID | FETCH-LOGICAL-c5382-841900fb1cfbcb62a4a976a2f68c1b39a08ff8553172a336eff96275b85a38b92 |
ISSN | 0006-3592 |
IngestDate | Tue Aug 27 13:48:48 EDT 2024 |
IsPeerReviewed | true |
IsScholarly | true |
Issue | 1 |
Language | English |
License | (c) 1993 John Wiley & Sons, Inc. |
LinkModel | OpenURL |
MergedId | FETCHMERGED-LOGICAL-c5382-841900fb1cfbcb62a4a976a2f68c1b39a08ff8553172a336eff96275b85a38b92 |
PMID | 18609651 |
ParticipantIDs | pubmed_primary_18609651 |
PublicationCentury | 1900 |
PublicationDate | 5 June 1993 |
PublicationDateYYYYMMDD | 1993-06-05 |
PublicationDate_xml | – month: 06 year: 1993 text: 5 June 1993 day: 05 |
PublicationDecade | 1990 |
PublicationPlace | United States |
PublicationPlace_xml | – name: United States |
PublicationTitle | Biotechnology and bioengineering |
PublicationTitleAlternate | Biotechnol Bioeng |
PublicationYear | 1993 |
SSID | ssj0007866 |
Score | 1.6735117 |
Snippet | Tyrosyl ring motions in alpha-lytic protease were investigated by solid-state deuterium nuclear magnetic resonance (NMR) spectroscopy in lyophilized enzyme... |
SourceID | pubmed |
SourceType | Index Database |
StartPage | 87 |
Title | Solid-state nuclear magnetic resonance investigation of solvent dependence of tyrosyl ring motion in an enzyme |
URI | https://www.ncbi.nlm.nih.gov/pubmed/18609651 |
Volume | 42 |
hasFullText | |
inHoldings | 1 |
isFullTextHit | |
isPrint | |
link | http://utb.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwnV1Jb9QwFLamIAQcEEzZF_mAuIxcJnHsOMe2AipQKyRaqbfK9tgoUsep2gCa_nqel0ySFsRyiSJ7Ennyvrwtz99D6LW1slRlpQgXbEEKm3EiFnpOpC8FKLNK64VP6O8f8L2j4uMxO55Mfgx3l7RqS1_-cl_J_0gVxkCufpfsP0h2fVMYgHOQLxxBwnD8Kxl_aU7rBQl7gmbOExPL89lSfnUmEjN7L9u_t3XPpRG9Q1iVL3OcdR1wdSwVWIHFXJ3OQkVe7O7jsyGgAIy7XI1JDXbqpl0n5cMHCFU3puc2XEf531L1z-c-afoh5Ili7N-39voEgbt0zfeorPb7bESs_eNkzkYalhPKqpGGLfJrSLoY2NprWjyywqq63YJoq_AuSj78HQjhbBlEmgnu2WuyP89eIdXupjbQRim8YjzwSZ5kwGGEx6gp_pVEzQqrejtakyecTfe5EpQE5-TwPrqXogq8HSHyAE2Mm6LNbSfbZrnCb3Co8w2ymqJbO93Z7d2u298U3R0QU24iN0AWTsjCHbLwGll4hCzcWJyQhXtk-dGELOxvjiOy4FIsHY7IeoiO3r873N0jqTMH0WAgwYQW4EfOrcq0VVrxXBYS3FqZWy50pmgl58JawUC_l7mklBtrfZMnpgSTVKgqf4RuuMaZJwiznJlMwfU5o0VpKwnxs6BUg3UwnGr6FD2OT_bkLNKvnHTP_NlvZ56jOz02X6CbFt538xKcx1a9CqL-CZaAcFQ |
link.rule.ids | 786 |
linkProvider | National Library of Medicine |
openUrl | ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fsummon.serialssolutions.com&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.genre=article&rft.atitle=Solid-state+nuclear+magnetic+resonance+investigation+of+solvent+dependence+of+tyrosyl+ring+motion+in+an+enzyme&rft.jtitle=Biotechnology+and+bioengineering&rft.au=Burke%2C+P+A&rft.au=Griffin%2C+R+G&rft.au=Klibanov%2C+A+M&rft.date=1993-06-05&rft.issn=0006-3592&rft.volume=42&rft.issue=1&rft.spage=87&rft_id=info:doi/10.1002%2Fbit.260420112&rft_id=info%3Apmid%2F18609651&rft_id=info%3Apmid%2F18609651&rft.externalDocID=18609651 |
thumbnail_l | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/lc.gif&issn=0006-3592&client=summon |
thumbnail_m | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/mc.gif&issn=0006-3592&client=summon |
thumbnail_s | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/sc.gif&issn=0006-3592&client=summon |