UniProt UniProt http://www.uniprot.org Alzheimer-associated cerebrospinal fluid fragments of neurogranin are generated by Calpain-1 and prolyl endopeptidase. Alzheimer-associated cerebrospinal fluid fragments of neurogranin are generated by Calpain-1 and prolyl endopeptidase. Molecular neurodegeneration 2018 imex curation Becker B., Nazir FH., Brinkmalm G., Camporesi E., Kvartsberg H., Portelius E., Boström M., Kalm M., Höglund K., Olsson M., Zetterberg H., Blennow K., Becker B., Nazir FH., Brinkmalm G., Camporesi E., Kvartsberg H., Portelius E., Boström M., Kalm M., Höglund K., Olsson M., Zetterberg H., Blennow K. bruno.becker@gu.se 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro protease assay protease assay ms participant Identification by mass spectrometry ms participant Becker B., Nazir FH., Brinkmalm G., Camporesi E., Kvartsberg H., Portelius E., Boström M., Kalm M., Höglund K., Olsson M., Zetterberg H., Blennow K., Becker B., Nazir FH., Brinkmalm G., Camporesi E., Kvartsberg H., Portelius E., Boström M., Kalm M., Höglund K., Olsson M., Zetterberg H., Blennow K. bruno.becker@gu.se Accepted 2020-FEB-27 AT 10:39 GMT AT 10:39 GMT by KALPANAP Molecular neurodegeneration 2018 Only protein-protein interactions imex curation imex curation Alzheimer-associated cerebrospinal fluid fragments of neurogranin are generated by Calpain-1 and prolyl endopeptidase. Alzheimer-associated cerebrospinal fluid fragments of neurogranin are generated by Calpain-1 and prolyl endopeptidase. Molecular neurodegeneration 2018 imex curation Becker B., Nazir FH., Brinkmalm G., Camporesi E., Kvartsberg H., Portelius E., Boström M., Kalm M., Höglund K., Olsson M., Zetterberg H., Blennow K., Becker B., Nazir FH., Brinkmalm G., Camporesi E., Kvartsberg H., Portelius E., Boström M., Kalm M., Höglund K., Olsson M., Zetterberg H., Blennow K. bruno.becker@gu.se 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro protease assay protease assay tag fluorescence tag visualisation by fluorescence tag fluorescence Becker B., Nazir FH., Brinkmalm G., Camporesi E., Kvartsberg H., Portelius E., Boström M., Kalm M., Höglund K., Olsson M., Zetterberg H., Blennow K., Becker B., Nazir FH., Brinkmalm G., Camporesi E., Kvartsberg H., Portelius E., Boström M., Kalm M., Höglund K., Olsson M., Zetterberg H., Blennow K. bruno.becker@gu.se Accepted 2020-FEB-27 AT 10:39 GMT AT 10:39 GMT by KALPANAP Molecular neurodegeneration 2018 Only protein-protein interactions imex curation imex curation Mock publication and experiment for abstract interactions that are not interaction evidences. Mock publication and experiment for abstract interactions that are not interaction evidences. unspecified method Alzheimer-associated cerebrospinal fluid fragments of neurogranin are generated by Calpain-1 and prolyl endopeptidase. Alzheimer-associated cerebrospinal fluid fragments of neurogranin are generated by Calpain-1 and prolyl endopeptidase. Molecular neurodegeneration 2018 imex curation Becker B., Nazir FH., Brinkmalm G., Camporesi E., Kvartsberg H., Portelius E., Boström M., Kalm M., Höglund K., Olsson M., Zetterberg H., Blennow K., Becker B., Nazir FH., Brinkmalm G., Camporesi E., Kvartsberg H., Portelius E., Boström M., Kalm M., Höglund K., Olsson M., Zetterberg H., Blennow K. bruno.becker@gu.se 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro protease assay protease assay western blot western blot Immuno blot Immuno blot Becker B., Nazir FH., Brinkmalm G., Camporesi E., Kvartsberg H., Portelius E., Boström M., Kalm M., Höglund K., Olsson M., Zetterberg H., Blennow K., Becker B., Nazir FH., Brinkmalm G., Camporesi E., Kvartsberg H., Portelius E., Boström M., Kalm M., Höglund K., Olsson M., Zetterberg H., Blennow K. bruno.becker@gu.se Accepted 2020-FEB-27 AT 10:39 GMT AT 10:39 GMT by KALPANAP Molecular neurodegeneration 2018 Only protein-protein interactions imex curation imex curation can1_human Calpain-1 catalytic subunit CAPN1 CANPL1 Calpain-1 large subunit Calcium-activated neutral proteinase 1 Calpain mu-type Micromolar-calpain Cell proliferation-inducing gene 30 protein PIG30 protein protein human Homo sapiens Human MSEEIITPVYCTGVSAQVQKQRARELGLGRHENAIKYLGQDYEQLRVRCLQSGTLFRDEAFPPVPQSLGYKDLGPNSSKTYGIKWKRPTELLSNPQFIVDGATRTDICQGALGDCWLLAAIASLTLNDTLLHRVVPHGQSFQNGYAGIFHFQLWQFGEWVDVVVDDLLPIKDGKLVFVHSAEGNEFWSALLEKAYAKVNGSYEALSGGSTSEGFEDFTGGVTEWYELRKAPSDLYQIILKALERGSLLGCSIDISSVLDMEAITFKKLVKGHAYSVTGAKQVNYRGQVVSLIRMRNPWGEVEWTGAWSDSSSEWNNVDPYERDQLRVKMEDGEFWMSFRDFMREFTRLEICNLTPDALKSRTIRKWNTTLYEGTWRRGSTAGGCRNYPATFWVNPQFKIRLDETDDPDDYGDRESGCSFVLALMQKHRRRERRFGRDMETIGFAVYEVPPELVGQPAVHLKRDFFLANASRARSEQFINLREVSTRFRLPPGEYVVVPSTFEPNKEGDFVLRFFSEKSAGTVELDDQIQANLPDEQVLSEEEIDENFKALFRQLAGEDMEISVKELRTILNRIISKHKDLRTKGFSLESCRSMVNLMDRDGNGKLGLVEFNILWNRIRNYLSIFRKFDLDKSGSMSAYEMRMAIESAGFKLNKKLYELIITRYSEPDLAVDFDNFVCCLVRLETMFRFFKTLDTDLDGVVTFDLFKWLQLTMFA 1CB6D7C56D063498 neug_mouse Neurogranin RC3 Nrgn protein protein mouse Mus musculus Mouse MDCCTESACSKPDDDILDIPLDDPGANAAAAKIQASFRGHMARKKIKSGECGRKGPGPGGPGGAGGARGGAGGGPSGD 8E47CDB38E095794 cpns1_human Calpain small subunit 1 Calcium-dependent protease small subunit 1 Calcium-dependent protease small subunit Calpain regulatory subunit Calcium-activated neutral proteinase small subunit CAPNS1 CAPN4 CAPNS protein protein human Homo sapiens Human MFLVNSFLKGGGGGGGGGGGLGGGLGNVLGGLISGAGGGGGGGGGGGGGGGGGGGGTAMRILGGVISAISEAAAQYNPEPPPPRTHYSNIEANESEEVRQFRRLFAQLAGDDMEVSATELMNILNKVVTRHPDLKTDGFGIDTCRSMVAVMDSDTTGKLGFEEFKYLWNNIKRWQAIYKQFDTDRSGTICSSELPGAFEAAGFHLNEHLYNMIIRRYSDESGNMDFDNFISCLVRLDAMFRAFKSLDKDGTGQIQVNIQEWLQLTMYS 17B87A8E47A90632 neug_human Neurogranin NRGN RC3 protein protein human Homo sapiens Human MDCCTENACSKPDDDILDIPLDDPGANAAAAKIQASFRGHMARKKIKSGERGRKGPGPGGPGGAGVARGGAGGGPSGD 0D6ED790A31D5785 ppce_mouse Prolyl endopeptidase Prep Pep Post-proline cleaving enzyme protein protein mouse Mus musculus Mouse MLSFQYPDVYRDETSVQEYHGHKICDPYSWLEDPDSEQTKAFVEAQNKITVPFLEQCPIRGLYKERMTELYDYPKYSCHFKKGKRYFYFYNTGLQNQRVLYVQDSLEGEARVFLDPNTLSDDGTVALRGYAFSEDGEYFAYGLSASGSDWVTIKFMKVDGAKELPDVLERVKFTCMAWTHDGKGMFYNSYPQQDGKSDGTETSTNLHQKLCYHVLGTDQSEDILCAEFPDEPKWMGGAELSDDGRYVLLSIWEGCDPVNRLWYCDLQQEPNGITGILKWVKLIDNFEGEYDYVTNEGTVFTFKTNRNSPNYRLINIDFTDPDESKWKVLVPEHEKDVLEWVACVRSNFLVLCYLHDVKNILQLHDLTTGALLKTFPLDVGSVVGYSGRKKDSEIFYQFTSFLSPGVIYHCDLTKEELEPMVFREVTVKGIDAADYQTIQIFYPSKDGTKIPMFIVHKKGIKLDGSHPAFLYGYGGFNISITPNYSVSRLIFVRHMGGVLAVANIRGGGEYGETWHKGGILANKQNCFDDFQCAAEYLIKEGYTSPKRLTINGGSNGGLLVAACANQRPDLFGCVIAQVGVMDMLKFHKFTIGHAWTTDYGCSDTKQHFEWLLKYSPLHNVKLPEADDIQYPSMLLLTADHDDRVVPLHSLKFIATLQYIVGRSRKQSNPLLIHVDTKAGHGAGKPTAKVIEEVSDMFAFIARCLNIEWIQ mint Cleaves peptide bonds on the C-terminal side of prolyl residues within peptides that are up to approximately 30 amino acids long 1B010D5D6CA73C0E capns1_capn1_human-nrgn-1 4 8 enzyme target enzyme target substrate substrate unspecified role unspecified role purified purified region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. 15 enzyme enzyme unspecified role unspecified role purified purified human-erythrocyte Homo sapiens erythrocyte Erythrocyte, Red blood cell; Corpuscle; RBC protein cleavage protein cleavage Fig. 3 capns1_capn1_human-nrgn-3 2 8 enzyme target enzyme target substrate substrate unspecified role unspecified role purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal fluorescent dye fluorescent dye label fluorescent dye n-term range n-terminal range n-term range n-term range n-terminal range n-term range chemical synthesis Chemical synthesis (Chemical synthesis) 15 enzyme enzyme unspecified role unspecified role purified purified human-erythrocyte Homo sapiens erythrocyte Erythrocyte, Red blood cell; Corpuscle; RBC protein cleavage protein cleavage Fig. 2 capns1_capn1_human-nrgn-4 2 8 enzyme target enzyme target substrate substrate unspecified role unspecified role purified purified n-terminal fluorescent dye fluorescent dye label fluorescent dye n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 15 enzyme enzyme unspecified role unspecified role purified purified human-erythrocyte Homo sapiens erythrocyte Erythrocyte, Red blood cell; Corpuscle; RBC protein cleavage protein cleavage Fig. 2 capns1_capn1_human-nrgn-2 4 8 enzyme target enzyme target substrate substrate unspecified role unspecified role purified purified ecoli Escherichia coli (strain K12) 15 enzyme enzyme unspecified role unspecified role purified purified human-erythrocyte Homo sapiens erythrocyte Erythrocyte, Red blood cell; Corpuscle; RBC protein cleavage protein cleavage Fig. 4 nrgn-prep-1 2 9 enzyme enzyme unspecified role unspecified role purified purified mouse-brain Mus musculus brain brain Brain [cerebrum] 6 enzyme target enzyme target substrate substrate unspecified role unspecified role purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal fluorescent dye fluorescent dye label fluorescent dye n-term range n-terminal range n-term range n-term range n-terminal range n-term range chemical synthesis Chemical synthesis (Chemical synthesis) protein cleavage protein cleavage To increase the chance of detecting a C-terminal-cleaving activity on Ng in such a complex matrix, mouse brain extract was initially size-separated, and the fractions were further analyzed for C-terminal peptide cleaving activity (quenched FRET peptide representing Ng70–78) Fig. 6 nrgn-prep-2 1 9 enzyme enzyme unspecified role unspecified role purified purified mouse-brain Mus musculus brain brain Brain [cerebrum] 8 enzyme target enzyme target substrate substrate unspecified role unspecified role purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal fluorescent dye fluorescent dye label fluorescent dye n-term range n-terminal range n-term range n-term range n-terminal range n-term range Q92686:p.Glu50delinsLysLysLysGlu mutation mutation certain certain sequence position certain certain certain sequence position certain E KKKE Tue Jun 30 19:25:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu50delinsLysLysLysGlu' chemical synthesis Chemical synthesis (Chemical synthesis) protein cleavage protein cleavage To increase the chance of detecting a C-terminal-cleaving activity on Ng in such a complex matrix, mouse brain extract was initially size-separated, and the fractions were further analyzed for C-terminal peptide cleaving activity (quenched FRET peptide representing Ng70–78) Fig. 9 capns1-capn1_human mu-Calpain complex CAPN1:CAPNS1 CAPN1/S1 complex Conventional Calpain complex CAPN1/CAPNS1 complex Calpain-1 complex 7 enzyme regulator enzyme regulator CAPN1 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 5 enzyme enzyme CAPNS1 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 43 41 physical association physical association human Homo sapiens Human stable complex stable complex biological system reconstruction evidence based on paralogy evidence used in manual assertion biological system reconstruction evidence based on paralogy evidence used in manual assertion Calpain (CAPN1/muCL) and its small regulatory subunit (CAPNS1/30K) bind via their respective 5th (of 5) EF-hand motifs. The N-terminus of CAPNS1 contains hydrophobic Gly-clusters, the Gly-rich domain. The catalytically-active cleft is rather deep and narrow which might explains calpain's preference for digesting inter-domain unstructured regions. It is formed by the two calpain protease core domains, PC1 and PC2, containing one Ca2+-binding sites per core. Calcium binding to several domains of both subunits releases the steric hindrance by allowing PC1 and PC2 to move closer to each other while the secondary structures remain essentially unchanged and may result in the dissociation of the small regulatory subunit from the catalytic subunits. Most other members of the calpain family of proteins do not require a regulatory subunit and are activated directly by calcium binding. Calpastatin (CAST, P20810) Calcium ion (Ca2+, CHEBI:29108) Muscular dystrophy [Orphanet:98473]: Muscular dystrophy (MD) refers to a group of more than 30 genetic diseases characterized by progressive weakness and degeneration of the skeletal muscles that control movement. Cataract [EFO:0001059]: Partial or complete opacity of the crystalline lens of one or both eyes that decreases visual acuity and eventually results in blindness. Some cataracts appear in infancy or in childhood, but most develop in older individuals. Alzheimer's disease [EFO:0000249]: A progressive, neurodegenerative disease characterized by loss of function and death of nerve cells in several areas of the brain leading to loss of cognitive function such as memory and language. Parkinson's disease [EFO:0002508]: A progressive degenerative disorder of the central nervous system characterized by loss of dopamine producing neurons in the substantia nigra and the presence of Lewy bodies in the substantia nigra and locus coeruleus. Signs and symptoms include tremor which is most pronounced during rest, muscle rigidity, slowing of the voluntary movements, a tendency to fall back, and a mask-like facial expression. Rheumatoid arthritis [EFO:0000685]: A chronic, systemic autoimmune disorder characterized by inflammation in the synovial membranes and articular surfaces. It manifests primarily as a symmetric, erosive polyarthritis that spares the axial skeleton and is typically associated with the presence in the serum of rheumatoid factor. Ischemia reperfusion injury [EFO:0002687]: Adverse functional, metabolic, or structural changes in ischemic tissues resulting from the restoration of blood flow to the tissue (reperfusion), including swelling; hemorrhage; necrosis; and damage from free radicals. The most common instance is myocardial reperfusion injury. A calcium-dependent protease complex that processes the substrate by limited proteolysis rather than degrading it. In some cases the proteolytic action activates the substrate, for example, it cleaves CDK5R1/p35 (Q15078) into its p25 form that is associated with Alzheimer's disease. Involved in cytoskeletal remodeling, signal transduction and implicated in cell cycle regulation and apoptosis. Finely-balanced calpain homeostasis is required as both over and under-activation causes disease. Calpain complexes recognise their substrates based on a short peptide sequence. Inhibited by the intrinsically-unstructured calpastatin (P20810) by its tight binding to the calpain catalytic subunit. In general, under disease/damaged conditions, such as muscular dystrophy, cardiomyopathy, traumatic ischemia, or lissencephaly, the over-activation of conventional calpains (probably due to compromised intracellular Ca2+ homeostasis) has been identified as an exacerbating factor. Autosomal recessive spastic paraplegia type 76 [EFO:0009019]: Spastic paraplegia-76 is an autosomal recessive neurologic disorder characterized by young-adult onset of slowly progressive spasticity of the lower limbs resulting in gait difficulties. Most affected individuals have upper limb involvement and additional features such as foot deformities and dysarthria. Cognition is unaffected. Accepted 2019-JUN-05 by ORCHARD