PDBsum entry 3wlc

Fluorescent protein

PDB id: 3wlc

Contents

Protein chain

393 a.a.

Metals

_CA ×4

Waters ×159

PDB id:

3wlc

Name:

Fluorescent protein

Title:

Crystal structure of dimeric gcamp6m

Structure:

Myosin light chain kinase, green fluorescent protein, calmodulin. Chain: a. Fragment: unp residues 37-55, 149-238, 2-144, 3-149. Synonym: cam. Engineered: yes. Mutation: yes. Other_details: this triple chimera comprises (from the n-terminus) an engineered expression tag, residues 37-55 of unp q6ldg3, an

Source:

Gallus gallus, aequorea victoria, rattus norvegicus. Chicken, jellyfish, rat. Organism_taxid: 9031, 6100, 10116. Gene: gfp, calm, cam. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

2.49Å R-factor: 0.179 R-free: 0.225

Authors:

J.Ding,A.F.Luo,L.Y.Hu,D.C.Wang,F.Shao

Key ref:

J.Ding et al. (2014). Structural basis of the ultrasensitive calcium indicator GCaMP6. Sci China Life Sci, 57, 269-274. PubMed id: 24390420 DOI: 10.1007/s11427-013-4599-5

Date:

08-Nov-13 Release date: 22-Jan-14

Protein chain

P0DP29  (CALM1_RAT) -  Calmodulin-1 from Rattus norvegicus

Seq: 149 a.a.

Struc: 393 a.a.*

Protein chain

P11799  (MYLK_CHICK) -  Myosin light chain kinase, smooth muscle from Gallus gallus

Seq: 1906 a.a.

Struc: 393 a.a.*

Protein chain

P42212  (GFP_AEQVI) -  Green fluorescent protein from Aequorea victoria

Seq: 238 a.a.

Struc: 393 a.a.*

* PDB and UniProt seqs differ at 263 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.2.7.11.18 - [myosin light-chain] kinase.
• Reaction:
  1. L-seryl-[myosin light chain] + ATP = O-phospho-L-seryl-[myosin light chain] + ADP + H⁺
  2. L-threonyl-[myosin light chain] + ATP = O-phospho-L-threonyl-[myosin light chain] + ADP + H⁺
• Cofactor: Ca(2+)

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

DOI no: 10.1007/s11427-013-4599-5

Sci China Life Sci 57:269-274 (2014)

PubMed id: 24390420

Structural basis of the ultrasensitive calcium indicator GCaMP6.

J.Ding, A.F.Luo, L.Hu, D.Wang, F.Shao.

ABSTRACT

GCaMP is one of the most widely used calcium indicators in neuronal imaging and calcium cell biology. The newly developed GCaMP6 shows superior brightness and ultrasensitivity to calcium concentration change. In this study, we determined crystal structures of Ca(2+)-bound GCaMP6 monomer and dimer and presented detailed structural analyses in comparison with its parent version GCaMP5G. Our analyses reveal the structural basis for the outperformance of this newly developed Ca(2+) indicator. Three substitution mutations and the resulting changes of local structure and interaction explain the ultrasensitivity and increased fluorescence intensity common to all three versions of GCaMP6. Each particular substitution in the three GCaMP6 is also structurally consistent with their differential sensitivity and intensity, maximizing the potential of using GCaMP6 in solving diverse problems in neuronal research and calcium signaling. Our studies shall also be beneficial to further structure-guided optimization of GCaMP and facilitate the design of novel calcium indicators.