Figure 3 of Jiang, Mol Vis 2005; 11:143-151.


Figure 3. The EF4 Ca2+ binding site and sequence alignment of vertebrate GCAPs

A: The Ca2+ binding loop consisting of 12 amino acids is flanked by two a-helical regions. Residues in the loop that coordinate the Ca2+ ion are marked with an asterisk. Although the 151 position amino acid (arrow) does not directly coordinate the Ca2+ ion, it is highly conserved throughout the GCAPs (leucine or isoleucine) suggesting it is essential for structure and function of this loop. B: The known vertebrate guanylate cyclase activating proteins (vertebrate GCAP1-8) have been aligned (only the C-terminal halves containing EF3 and EF 4 are shown). Residues conserved in more than 50% of the sequences shown are red. Conservative substitutions are blue (for origin of sequences and accession numbers, see Methods).

(18 K)

B:

                                                                               L151F
                                                                      I143NT     |  E155G
                   Y99C                                                  |       |   |
                    |    EF3                                             |  EF4  |   |
                    |------------                                        |------------
hGCAP1      94 WYFKLYDVDGNGCIDRDELLTIIQAIRAINPCSDTTMTAE---------EFTDTVFSKIDVNGDGELSLEEFIEGVQKDQMLLDTLTRSLDLTRIVRRLQNGEQDEEGADEAAEAAG
bGCAP1      94 WYFKLYDVDGNGCIIRDELLTIIRAIRAINPCSDSTMTAE---------EFTDTVFSKIDVNGDGELSLEEFMEGVQKDQMLLDTLTRSLDLTRIVRRLQNGEQDEEGASGRETEAAEADG
mGCAP1      94 WYFKLYDVDGNGCIDRDELLTIIRAIRTINPWSDSSMSAE---------EFTDTVFAKIDINGDGELSLEEFMEGVQKDQMLLDTLTRSLDLTGIVRRLQNGEHEEAGTSDLAAEAAG
ratGCAP1    94 WYFKLYDVDGNGCIDRDELLTIIRAIRTINPWSDSSMSAE---------EFTDTVFAKIDINGDGELSLEEFMEGVQKDQMLLDTLTRSLDLTRIVRRLQNGEQEEAGAGDLAAEAAG
fGCAP1      93 WYFKLYDVDGNGCIDRGELLNIIKAIRAINRCN-DEMTAE---------EFTDMVFDKIDINGDGELSLEEFIEGVQKDELLLEVLTRSLDLKHIVYMIQNDGKRMEISERPRQEITTGNSLP
cGCAP1      93 WYFKLYDVDGNGCIDRGELLNIIKAIRAINRCN-EAMTAE---------EFTNMVFDKIDINGDGELSLEEFMEGVQKDEVLLDILTRSLDLTHIVKLIQNDGKNPHAPEEAEEAAQ
oryGCAP1    93 WYFKLYDVDGNGCIDRHELLNIIKAIRAINGNENQEMTAE---------EFTNNVFDRIDVNGDGELSLEEFVEGARSDEDFMEVMMKSLDLRHIVAMIHNRRHSV
fuguGCAP1   93 WYFKLYDVDGNGCIDRHELLNIIKAIRAINGNENHETTAE---------DFTNSVFDRIDINGDGELSLEEFVAGARSDDVFMEVMIKSLDLTHIVAMIHNRRHSV
zGCAP1      93 WYFRLYDVDGNGCIDRYELLNIIKAIRAINGSETQESSAE---------EFTNRVFERIDINGDGELSLDEFVAGARSDEEFMEAMMKSLDLTHIVAMIHNRRHSV
bGCAP2      99 WTFKIYDKDRNGCIDRQELLDIVESIYKLKKACSVEVEAEQQGKLLTPEEVVDRIFLLVDENGDGQLSLNEFVEGARRDKWVMKMLQ--MDLNPSSWISQQRRKSAMF
hGCAP2      96 WTFKIYDKDGNGCIDRLELLNIVEGIYQLKKACRRELQTEQD-QLLTPEEVVDRIFLLVDENGDGQLSLNEFVEGARRDKWVMKMLQ--MDMNPSSWLAQQRRKSAMF
mGCAP2      96 WTFKIYDKDRNGCIDRLELLDIVEAIYKLKKACRAELDLEHQGQLLTPEEVVDRIFLLVDENGDGQLSLTEFIEGARRDKWVMKMLQ--MDINPGGWITQQRRRSAMF
fuguGCAP2   95 WTFKMYDKDGSGCIDKTELLEIVESIYRLKKACHGELDEDCT--LLTPDQVVDRIFELVDENGDGELSLDEFIDGARRDKWVMKMLQ--MDVNPGDWLND-RRRSADF
oryGCAP2    95 WTFKMYDKDGSGCIDKTELLEIVESIYRLKKACHGELDEECN--LLTPDQVVDRIFELVDENGDGELSLDEFIDGARRDKWVMKMLQ--MDVNPGDWINE-RRCSEDF
zGCAP2      94 WTFKMYDKDGSGCIDKTELKEIVESIYRLKKACHGELDAECN--LLTPDQVVARIFELVDENGDGELSLDEFIAGARRDKWVMKMLQ--MDVNPGDWINEQRRRSANF
fGCAP2      94 WTFKVYDRDGNGCIDKTELLEIVESIYNLKKVCRQGQDDRIP--LLSPEQVVDRIFQLVDENGDGQLSLDEFIDGARKDKWVMKMLQ--MDVSPGSWINEQRRKSALF
cGCAP2      95 WTFKVYDKDGNGCIDKPELLEIVESIYKLKKVCRSEVEERTP--LLTPEEVVDRIFQLVDENGDGQLSLDEFIDGARKDKWVMKMLQ--MDVNPGGWISEQRRKSALF
siluGCAP2   92 WSFKIYDKDGNGCVDKRELKEIIQSIYSIKRGWRR--DQEAQ--LMSPEEICERIFQIVDENGDGQLSLQEFVEGAXKDTWVLKMLQ--LDTNPCNWVMEQRRKSALF
fuguGCAP6   93 WSFKVYDRDNNGFVDRTELRSIIDSIHRIKK------DSGSQ---LTVDEVVDRIFQAADSDGDGYISAEEFIRGAQQDPWLLNILK--LDMNPAGWVMEQRRKSAHF
fuguGCAP7   92 WSFKMYDKDGNGKLDRQEVKRLIKIIHKIKLQRSDINMTP--------SEICDRIFELVDNNNDGQISLSEFMEGAQKDEWVMNLLK--LDVNATSWVLHNCEKLP
fuguGCAP8   94 WSFKVYDRDGNGCLDKQEVRHIVKIIHKIK-KKSDMSVTEKI------EDVCDRIFELVDKNKDSQISLEEFIEGAEKDPWLMNQLR--LDIGPCEWFIEQREKKL
fuguGCAP5   92 WYFKLYDIDGSGCIDRDELLLIIKSIRAINGIP-SEMSAE---------EFTNMVFDKIDINGDGELSYEEFIEGIQNDETLLKMLTESLDLTHIMQKIEGQMNASSSD
zGCAP5      93 WYFKLFDMDGSGCIDKDELLLIFKAVQAINGAE-PEISAE---------DLADMVFNKIDVNGDGELSLEEFMEGISADEKISEMLTQSLDLTRIVSNIYNDSYIEQEAEIIEDQA
zGCAP3      92 WYFKLFDQDGNGKIDRDEMETIFKAIQDITRSY--EIPPD---------DIVSLIYERIDVNNEGELTLEEFITGAKEHPDIMEMLTKMMDLTHVLEIIVNGQKKKKE
fuguGCAP3   92 WYFKLFDQDGNGKIDKDELETIFKAIQDITRTY--DIPPE---------EIVTLIYDKIDVNGEGELTLEEFISGAKEHPDIMEMLTKMMDLSHVLEIIIKGQKKNTVK
zGCAP4      92 WYFKLFDQDGNGKIDKDELETIFTAIQDITRNR--DIVPE---------EIVALIFEKIDVNGEGELTLEEFIEGAKEHPEIMDMLKILMDLTPVLLIIVEGRQK
fuguGCAP4   92 WYFKLFDQDGNGKIDREELETIFSAIQDITRNR--DIDPE---------EIVSLIFERIDVNGEGELTLEEFIEGAKDHPDIMDTLKKIMDLTPVLVIIVEGRTG
hGCAP3      95 WYFKLYDADGNGSIDKNELLDMFMAVQALNGQQ--TLSPE---------EFINLVFHKIDINNDGELTLEEFINGMAKDQDLLEIVYKSFDFSNVLRVICNGKQPDMETDSSKSPDKAGLGKVKMK
fGCIP       94 WSFKLYDKDGDGAITRSEMLEIMRAVYKMSVVASLTKVNPMT-----AEECTNRIFVRLDKDQNAIISLQEFVDGSLGDEWVRQMLE--CDLSTVEIQKMTKHSHLPARSSRERLFHANT
fuguGCIP    95 WSFKLYDKDRDGGITRQEMLEIMQAVYKMSLAAALTRPNPLT-----AEECTNRVFARLDRDNNAIISLEEFIEGALDDDWIREMLE--CDPTTVKVERPLRRDAVLGIHG
                     ------------                                         ------------
                         EF3                                                  EF4

Jiang, Mol Vis 2005; 11:143-151 <http://www.molvis.org/molvis/v11/a16/>
©2005 Molecular Vision <http://www.molvis.org/molvis/>
ISSN 1090-0535