Functional analysis of the guanylate kinase-like domain in the synapse-associated protein SAP97.
SAP97 is a membrane cytoskeletal protein localized at the presynaptic nerve terminals of type 1 asymmetric synapses. It has been implicated in the assembly of synapses and in particular in the localization and clustering of ion channels. The C-terminal GK domain of SAP97 shares a high degree of sequence similarity with low-molecular-mass guanylate kinases. These enzymes are involved in the guanine nucleotide metabolic cycle and in the maintenance of GTP/GDP pools required for example in Ras-mediated cell signaling. It has therefore been hypothesized that SAP97 plays an essential role in cellular signaling by regulating the guanine nucleotide pools at synaptic junctions. Here, we test this hypothesis by assessing whether the GK domain in SAP97 encodes an authentic guanylate kinase. We show that the GK domain in and of itself does not encode an active guanylate kinase, that it cannot be activated by its binding partner GKAP and that flanking regions are not acting as inhibitory regulators for enzymatic activity. Thus, it would appear that the GK domain of SAP97 is not involved in the metabolism of guanine nucleotides required for signaling events.
Kuhlendahl
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Department of Neurobiology, University of Alabama at Birmingham, 35294-0021, USA.
Spangenberg
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Konrad
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Kim
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E
Garner
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CC
D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002352:Carrier Proteins / Q000494:pharmacology; D005816:Genetic Complementation Test; D006150:Guanine Nucleotides / Q000502:physiology; D051528:Guanylate Kinases; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000737:chemistry*; D009703:Nucleoside-Phosphate Kinase / Q000737:chemistry*; D011485:Protein Binding / Q000502:physiology; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D000074407:SAP90-PSD95 Associated Proteins; D016415:Sequence Alignment
D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; C501119:Dlg1 protein, rat; D006150:Guanine Nucleotides; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D011993:Recombinant Fusion Proteins; D000074407:SAP90-PSD95 Associated Proteins; D009703:Nucleoside-Phosphate Kinase; D051528:Guanylate Kinases