BMRB Entry 30936

Title:
Atomic-Resolution Structure of Kinesin-1 Motor Domain in Complex with Polymeric Microtubules by Magic Angle Spinning NMR
Deposition date:
2021-07-20
Original release date:
2022-11-17
Authors:
Zhang, C.; Guo, C.; Russell, R.; Quinn, C.; Li, M.; Williams, J.; Gronenborn, A.; Polenova, T.
Citation:

Citation: Zhang, C.; Guo, C.; Russell, R.; Quinn, C.; Li, M.; Williams, J.; Gronenborn, A.; Polenova, T.. "Magic-angle-spinning NMR structure of the kinesin-1 motor domain assembled with microtubules reveals the elusive neck linker orientation"  Nat. Commun. 13, 6795-6795 (2022).
PubMed: 36357375

Assembly members:

Assembly members:
entity_1, polymer, 349 residues, 39318.406 Da.

Natural source:

Natural source:   Common Name: Human   Taxonomy ID: 9606   Superkingdom: Eukaryota   Kingdom: Metazoa   Genus/species: Homo sapiens

Experimental source:

Experimental source:   Production method: recombinant technology   Host organism: Escherichia coli

Data sets:
Data typeCount
13C chemical shifts1098
15N chemical shifts278

Additional metadata:

  • Assembly
  • Samples and Experiments
  • Software
  • Spectrometers
  • Hide all

Assembly:

Entity Assembly IDEntity NameEntity ID
1unit_11

Entities:

Entity 1, unit_1 349 residues - 39318.406 Da.

1   METALAASPLEUALAGLUCYSASNILELYS
2   VALMETCYSARGPHEARGPROLEUASNGLU
3   SERGLUVALASNARGGLYASPLYSTYRILE
4   ALALYSPHEGLNGLYGLUASPTHRVALVAL
5   ILEALASERLYSPROTYRALAPHEASPARG
6   VALPHEGLNSERSERTHRSERGLNGLUGLN
7   VALTYRASNASPCYSALALYSLYSILEVAL
8   LYSASPVALLEUGLUGLYTYRASNGLYTHR
9   ILEPHEALATYRGLYGLNTHRSERSERGLY
10   LYSTHRHISTHRMETGLUGLYLYSLEUHIS
11   ASPPROGLUGLYMETGLYILEILEPROARG
12   ILEVALGLNASPILEPHEASNTYRILETYR
13   SERMETASPGLUASNLEUGLUPHEHISILE
14   LYSVALSERTYRPHEGLUILETYRLEUASP
15   LYSILEARGASPLEULEUASPVALSERLYS
16   THRASNLEUSERVALHISGLUASPLYSASN
17   ARGVALPROTYRVALLYSGLYCYSTHRGLU
18   ARGPHEVALCYSSERPROASPGLUVALMET
19   ASPTHRILEASPGLUGLYLYSSERASNARG
20   HISVALALAVALTHRASNMETASNGLUHIS
21   SERSERARGSERHISSERILEPHELEUILE
22   ASNVALLYSGLNGLUASNTHRGLNTHRGLU
23   GLNLYSLEUSERGLYLYSLEUTYRLEUVAL
24   ASPLEUALAGLYSERGLULYSVALSERLYS
25   THRGLYALAGLUGLYALAVALLEUASPGLU
26   ALALYSASNILEASNLYSSERLEUSERALA
27   LEUGLYASNVALILESERALALEUALAGLU
28   GLYSERTHRTYRVALPROTYRARGASPSER
29   LYSMETTHRARGILELEUGLNASPSERLEU
30   GLYGLYASNCYSARGTHRTHRILEVALILE
31   CYSCYSSERPROSERSERTYRASNGLUSER
32   GLUTHRLYSSERTHRLEULEUPHEGLYGLN
33   ARGALALYSTHRILELYSASNTHRVALCYS
34   VALASNVALGLULEUTHRALAGLUGLNTRP
35   LYSLYSLYSTYRGLULYSGLULYSGLU

Samples:

sample_1: Apo-Kif5b, [U-100% 13C; U-100% 15N], 9.1 % w/w; Porcine-alpha-tubulin 16.4 % w/w; Porcine-beta-tubulin 16.4 % w/w; TAXOL 5 uM; GUANOSINE-5'-TRIPHOSPHATE 1 mM; GUANOSINE-5'-DIPHOSPHATE 1 mM; Buffer 58.1 % w/w

sample_2: Apo-Kif5b, [U-100% 13C; U-100% 15N], 9.1 % w/w; Porcine-alpha-tubulin 16.4 % w/w; Porcine-beta-tubulin 16.4 % w/w; TAXOL 5 uM; GUANOSINE-5'-TRIPHOSPHATE 1 mM; GUANOSINE-5'-DIPHOSPHATE 1 mM; Buffer 58.1 % w/w

sample_3: Apo-Kif5b, [1,6-13C-glucose; U-100% 15N], 8.2 % w/w; Porcine-alpha-tubulin 16.4 % w/w; Porcine-beta-tubulin 16.4 % w/w; TAXOL 5 uM; GUANOSINE-5'-TRIPHOSPHATE 1 mM; GUANOSINE-5'-DIPHOSPHATE 1 mM; Buffer 59 % w/w

sample_4: Apo-Kif5b, [2-13C-glucose; U-100% 15N], 7.6 % w/w; Porcine-alpha-tubulin 16.4 % w/w; Porcine-beta-tubulin 16.4 % w/w; TAXOL 5 uM; GUANOSINE-5'-TRIPHOSPHATE 1 mM; GUANOSINE-5'-DIPHOSPHATE 1 mM; Buffer 59.6 % w/w

sample_conditions_1: ionic strength: 100 mM; pH: 7.4; pressure: 1 atm; temperature: 283 K

Experiments:

NameSampleSample stateSample conditions
2D 13C-detected CORDsample_1isotropicsample_conditions_1
2D 13C-detected CORDsample_2isotropicsample_conditions_1
2D 13C-detected CORDsample_3isotropicsample_conditions_1
2D 13C-detected CORDsample_4isotropicsample_conditions_1
2D NCACXsample_2isotropicsample_conditions_1
3D NCOCXsample_2isotropicsample_conditions_1
3D NCACXsample_2isotropicsample_conditions_1

Software:

TopSpin, Bruker Biospin - collection, processing

NMRPipe, Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax - processing

CcpNmr Analysis, CCPN - chemical shift assignment, peak picking

TALOS-N, Cornilescu, Delaglio and Bax - chemical shift calculation

X-PLOR NIH, Schwieters, Kuszewski, Tjandra and Clore - refinement, structure calculation

NMR spectrometers:

  • Bruker AVANCE III 850 MHz
  • Bruker AVANCE NEO 600 MHz