University of Groningen

Post-translational modification of therapeutic peptides by NisB, the dehydratase of thelantibiotic nisinKluskens, LD; Kuipers, A; Rink, R; de Boef, E; Fekken, S; Driessen, Arnold; Kuipers, Oscar;Moll, GertPublished in:Biochemistry

DOI:10.1021/bi050805p

IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite fromit. Please check the document version below.

Document VersionPublisher's PDF, also known as Version of record

Publication date:2005

Link to publication in University of Groningen/UMCG research database

Citation for published version (APA):Kluskens, L. D., Kuipers, A., Rink, R., de Boef, E., Fekken, S., Driessen, A. J. M., ... Moll, G. N. (2005).Post-translational modification of therapeutic peptides by NisB, the dehydratase of the lantibiotic nisin.Biochemistry, 44(38), 12827-12834. DOI: 10.1021/bi050805p

CopyrightOther than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of theauthor(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).

Take-down policyIf you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediatelyand investigate your claim.

Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons thenumber of authors shown on this cover page is limited to 10 maximum.

Download date: 10-02-2018

http://dx.doi.org/10.1021/bi050805phttps://www.rug.nl/research/portal/en/publications/posttranslational-modification-of-therapeutic-peptides-by-nisb-the-dehydratase-of-the-lantibiotic-nisin(5fde9f66-c503-4f6d-8f89-6643363133b2).html

Post-translational modification of therapeutic peptides by NisB, the dehydratase of the lantibiotic nisin

Leon D. Kluskens, Anneke Kuipers, Rick Rink, Esther de Boef, Susan Fekken,

Arnold J.M. Driessen, Oscar P. Kuipers and Gert N. Moll

Supporting information Figure S1

3339 3678 4017

Mass (m/z)

3581.8

3600.03714.7

3730.7

Mal

di T

OF

sign

al

%

100

Figure S2

3360.4 3578.8 3797.2

Mass (m/z)

3570.7

3552.3100

Mal

di T

OF

sign

al

%3688.6

Figure S3

3438.6 3586.2 3733.8

Mass (m/z)

3570.6

Mal

di T

OF

sign

al

100

%3689.8

3702.0

Figure S4

3264.8

3200 3300

Mass (m/z)

Mal

di T

OF

sign

al100

%

3297.7

Figure S5

3120.0 3300.4 3480.8

100

Mass (m/z)

3327.6

3345.3

Mal

di T

OF

sign

al

%

Figure S6

3298 3444

100

3283

.8

3302

.4

3396

.6

3320

.1

3432

.7

3450

.7

3338

.7

3495

.3

3265

.5

Mass (m/z)

Mal

di T

OF

sign

al

Figure S7

Mass (m/z)

Mal

di T

OF

sign

al

3503.4 3559.6 3615.8

3518.2

3500.3100

%

Figure S8

3506.8

3488.9

Mass (M/z)

3499 3620 3741

Mal

di T

OF

sign

al

100

%

3637.5

Figure S9

Mal

di T

OF

sign

al5499.8

3799.8 5200.2 6600.6

100

%

Mass (m/z)

5644.4

Figure S10

3135.7 3307.5

3176

.0

3157.9100

Mal

di T

OF

sign

al

Mass (m/z)

%

Figure S11

3536.8 4051.6

1003784.1

Mass (m/z)

Mal

di T

OF

sign

al

%

3915.73815.9

Figure S12

LHRH: 1172.6 Da

Retention time (min)

20 30 40

24.02

mA

U

Legend Figure S1-11. Maldi-TOF mass spectrometry of NisB-dehydrated therapeutic peptides that are attached to the nisin leader peptide without the starting methionine. Figure S1: ACTH(1-10): leader-SYSMECFRWG; Figure S2: S1G-ACTH(1-10): leader-GYSMECFRWG; Figure S3: S3G-ACTH(1-10): leader-SYGMECFRWG; Figure S4: VP1: leader-ATFQCAPRG, Figure S5: VP2: leader-AYTQNCPRG; Figure S6: TPPII: leader-ITSISRASVA, Figure S7: enkA: leader-IAAIRYTGFC, Figure S8: enkB, leader-IAAIRYSGFC; Figure S9: LHRH: leader-ITSISLCTPGCKTGALMGCNKQHWSYGCRPG. Figure S10: Ang(1-7): leader-DRVTIHC; Figure S11: EPO(1-13): leader-YASHFGPLGWVCK. Figure S12. Reversed-phase HPLC UV absorbance chromatogram of trypsin-digested leader-LHRH, measured at 280 nm. The dominant LHRH peak (QHWSYGCRPG) was collected and confirmed by Maldi-TOF mass spectrometry.