Crystal Structure of G Protein-Coupled Receptor Kinase 5 (GRK5) in Complex with Calmodulin (CaM)

Structure of a GRK5-Calmodulin Complex Reveals Molecular Mechanism of GRK Activation and Substrate Targeting.

(ACE)MELENIVANTVLLKAREGGGGKRKGKSKKWKEILKFPHISQCEDLRRTIDRDYCSLCDKQPIGRLLFRQFCETRP
GLECYIQFLDSVAEYEVTPDEKLGEKGKKIMTKYLTPKSPVFIAQVGQDLVSQTEEKLLQKPCKELFSACAQSVHEYLRG
EPFHEYLDSMFFDRFLQWKWLERQPVTKNTFRQYRVLGKGGFGEVCACQVRATGKMYACKRLEKKRIKKRKGESMALNEK
QILEKVNSQFVVNLAYAYETKDALCLVLTIMNGGDLKFHIYNMGNPGFEEERALFYAAEILCGLEDLHHENTVYRDLKPE
NILLDDYGHIRISDLGLAVKIPEGDLIRGRVGTVGYMAPEVLNNQRYGLSPDYWGLGCLIYEMIEGQSPFRGRKEKVKRE
EVDRRVLETEEVYSHKFSEEAKSICKMLLTKDAKQRLGCQEEEAAEVKRHPFFRNMNFKRLEAGMLDPPFVPDPRAVYCK
DVLDIEQFSTVKGVNLDHTDDDFYSKFSTGSVSIPWQNEMIETECFKELNVFGPNGTLPPDLNRNHPPEPPKKGLLQRLF
KRQHQNNSKSSPSSKTSFNHHINSNHVSSNSTGSS

XMELENIVANTVLLKAREGGGGKRKGKSKKWKEILKFPHISQCEDLRRTIDRDYCSLCDKQPIGRLLFRQFCETRPGLEC
YIQFLDSVAEYEVTPDEKLGEKGKKIMTKYLTPKSPVFIAQVGQDLVSQTEEKLLQKPCKELFSACAQSVHEYLRGEPFH
EYLDSMFFDRFLQWKWLERQPVTKNTFRQYRVLGKGGFGEVCACQVRATGKMYACKRLEKKRIKKRKGESMALNEKQILE
KVNSQFVVNLAYAYETKDALCLVLTIMNGGDLKFHIYNMGNPGFEEERALFYAAEILCGLEDLHHENTVYRDLKPENILL
DDYGHIRISDLGLAVKIPEGDLIRGRVGTVGYMAPEVLNNQRYGLSPDYWGLGCLIYEMIEGQSPFRGRKEKVKREEVDR
RVLETEEVYSHKFSEEAKSICKMLLTKDAKQRLGCQEEEAAEVKRHPFFRNMNFKRLEAGMLDPPFVPDPRAVYCKDVLD
IEQFSTVKGVNLDHTDDDFYSKFSTGSVSIPWQNEMIETECFKELNVFGPNGTLPPDLNRNHPPEPPKKGLLQRLFKRQH
QNNSKSSPSSKTSFNHHINSNHVSSNSTGSS

ADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTD
SEEEIREAFRVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK

Sample- 25 mg/ml GRK5/CaM complex in buffer 20 mM Hepes, pH 7.2, 200 mM NaCl, 2 mM CaCl2, 1 mM DTT incubated with SGV in the presence of MgCl2.

Crystallization screening reservoir condition- 90% of (200 mM KCl, 50 mM Hepes pH 7.5, 30% 5/4 PO/OH) + 10% of (0.05% w/v L-Citrulline, 0.05% w/v Glycine, 0.05% w/v L-(-)-Threonine, 0.05% w/v L-(+)-Lysine, 0.05% w/v L-Alanine, 0.05% w/v L-Arginine, 0.05% w/v L-Asparagine monohydrate, 0.05% w/v L-Aspartic acid, 0.05% w/v L-Glutamic acid, 0.05% w/v L-Glutamine, 0.05% w/v L-Histidine, 0.05% w/v L-Isoleucine, 0.05% w/v L-Leucine, 0.05% w/v L-Methionine, 0.05% w/v L-Phenylalanine, 0.05% w/v L-Proline, 0.05% w/v L-Serine, 0.05% w/v L-Tryptophan, 0.05% w/v L-Tyrosine, 0.05% w/v L-Valine, 0.02 M HEPES sodium pH 6)

National Institutes of Health/National Heart, Lung, and Blood Institute (NIH/NHLBI)

National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)

4tnb contains same protein complexed with Sangivamycicn only

4tnD contains same protein complexed with AMP-PNP

G protein-coupled receptor (GPCR); G protein-coupled receptor kinase; phosphorylation; protein kinase; receptor regulation; signal transduction; Calmodulin; Calcium binding protein, TRANSFERASE

MELENIVANTVLLKAREGGGGKRKGKSKKWKEILKFPHISQCEDLRRTIDRDYCSLCDKQPIGRLLFRQFCETRPGLECY
IQFLDSVAEYEVTPDEKLGEKGKEIMTKYLTPKSPVFIAQVGQDLVSQTEEKLLQKPCKELFSACAQSVHEYLRGEPFHE
YLDSMFFDRFLQWKWLERQPVTKNTFRQYRVLGKGGFGEVCACQVRATGKMYACKRLEKKRIKKRKGESMALNEKQILEK
VNSQFVVNLAYAYETKDALCLVLTIMNGGDLKFHIYNMGNPGFEEERALFYAAEILCGLEDLHRENTVYRDLKPENILLD
DYGHIRISDLGLAVKIPEGDLIRGRVGTVGYMAPEVLNNQRYGLSPDYWGLGCLIYEMIEGQSPFRGRKEKVKREEVDRR
VLETEEVYSHKFSEEAKSICKMLLTKDAKQRLGCQEEGAAEVKRHPFFRNMNFKRLEAGMLDPPFVPDPRAVYCKDVLDI
EQFSTVKGVNLDHTDDDFYSKFSTGSVSIPWQNEMIETECFKELNVFGPNGTLPPDLNRNHPPEPPKKGLLQRLFKRQHQ
NNSKSSPSSKTSFNHHINSNHVSSNSTGSS