Region Pre-S1
Ala1 - conserved - Ser, Ala
- alpha-helix H1
Thr2 - conserved - Ala
- alpha-helix H1
Leu3 - conserved - Ile
- alpha-helix H1
Asp4 - not conserved - Glu, Ser
- alpha-helix H1
Ser5 - conserved - Glu
- alpha-helix H1
Trp6 - conserved
- not protected by acarbose N-bromosuccinimide
- alpha-helix H1
Leu7 - conserved - Ile
- alpha-helix H1
Ser8 - not conserved - Lys, Gln
- alpha-helix H1
Asn9 - not conserved - Lys, Arg
- alpha-helix H1
Glu10 - conserved - Gln, Glu
- not protected by acarbose EAC
- alpha-helix H1
Ala11 - not conserved - Glu, Lys
- alpha-helix H1
Thr12 - not conserved - Gly, Lys
- alpha-helix H1
Val13 - conserved - Ile
- alpha-helix H1
Ala14 - conserved - Ser
- alpha-helix H1
Arg15 - conserved - Ile
- alpha-helix H1
Thr16 - not conserved - Phe, Glu
- alpha-helix H1
Ala17 - conserved - Arg
- alpha-helix H1
Ile18 - conserved - Met
- alpha-helix H1
Leu19 - conserved - Phe
- alpha-helix H1
Asn20 - not conserved - Glu, Gln, Arg
- alpha-helix H1
Asn21 - conserved
- alpha-helix H1
Ile22 - conserved
- alpha-helix H1
Gly23 - conserved - Ala
- alpha-helix H1
Ala24 - not conserved - Tyr
Asp25 - conserved - Pro, Asp
- non-reacting group EAC
Gly26 - not conserved - Ser, Glu
Ala27 - conserved - Gly
Trp28 - not conserved - Val, Gln, Asn
- not protected by acarbose N-bromosuccinimide
Val29 - not conserved - Tyr, Phe, Pro
Ser30 - conserved - Pro, Ser, Asn
Gly31 - not conserved - Ser, Gly, Asn
Ala32 - not conserved - Ile, Gly, Ser
Asp33 - not conserved - Val, Leu, Ser, Ala
- non-reacting group EAC
Ser34 - conserved - Pro, Thr, Ser
Region S1
Gly35 - conserved
Ile36 - not conserved - Thr, Val, Phe
Val37 - conserved - Ile
Val38 - not conserved - Ile , Ala
Ala39 - conserved
...Sf... *Val39 - increase hydrophobicity
- same activity at 30deg. C
- no activity at 50deg. C
- no volumous groups accepted
...Sf... *Asp39 - decrease hydrophobicity
- no activity at 30deg. or 50deg. C
- no charged groups accepted
Ser40 - conserved
Pro41 - conserved - Leu
Ser42 - conserved
- H-bond carbonyl-O Asp44
Thr43 - conserved - Glu
Asp44 - not conserved - Ser, Thr, Ala
- non-reacting group EAC
- H-bond with sugar Asn395
Asn45 - not conserved - His, Gly, Asn
Pro46 - conserved
Asp47 - conserved
- not protected by acarbose EAC
- H-bond (3.4 Å) carbonyl-O Thr43
- H-bond (3.4 Å) Tyr95
- H-bond amide-N Phe49
...Sf... *Tyr47 - study of carboxyl group in S1
- no activity at 30deg. or 50deg. C
Tyr48 - conserved
- non-reacting group EAC
- H-bond Glu400
- H-bond Tyr311
- H-bond amide-N Phe48 ??
Phe49 - conserved - Tyr
Tyr50 - conserved
- non-reacting group EAC
- near Trp120
- H-bond (3.4 Å) carbonyl-O Asp47 ??
Thr51 - not conserved - Glu, Ala
Trp52 - conserved
- non-reacting group N-bromosuccinimide
- H-bond Glu179 (w/ Trp120 , Gln124)
- near Arg54, Asp176, ??
...Sf... *Arg52 - no activity at 30deg. or 50deg. C
...Sf... *Leu52 - no activity at 30deg. or 50deg. C
Thr53 - conserved - Ile
Arg54 - conserved
- H-bond Asp126
- H-bond Gln124
- H-bond Asp126 ??
- near Trp52 , Asp179
- alpha-helix H82
...Sf... *Lys54 - study of another basic residue
- no activity at 30deg. or 50deg. C
...Sf... *Thr54 - study of a polar but uncharged residue
- no activity at 30deg. or 50deg. C
Asp55 - conserved
- not protected by acarbose EAC
- cleft near Trp417
- interaction with Thr416 on free side of chain in active site ??
- alpha-helix H82
*Gly55 - decrease 22x in ko
- same Km
- selectivity data not available yet
...Sf... *Asn55 - no activity at 30deg. or 50deg. C
...Sf... *Tyr55 - no activity at 30deg. or 50deg. C
Ser56 - conserved - Ala
- alpha-helix H82
Gly57 - conserved - Ala , Gly
- alpha-helix H82
Leu58 - conserved - Ile
- alpha-helix H82
Val59 - conserved - Thr, Val
- alpha-helix H82
Region Inter S1 - S2
Leu60 - not conserved - Ile, Phe, Ser
- alpha-helix H82
Lys61 - conserved - Asn, Leu, Lys
- alpha-helix H82
Thr62 - not conserved - Thr, Ser, Val
- alpha-helix H82
- H-bond Asp65
Leu63 - conserved - Ile, Val, Leu
- alpha-helix H82
Val64 - conserved - Leu
- alpha-helix H82
Asp65 - not matched
- not protected by acarbose EAC
- alpha-helix H82
- H-bond Lys61
Leu66 - conserved - Ser, Tyr, Leu
- alpha-helix H82
Phe67 - not conserved - Glu, His
- alpha-helix H82
Arg68 - not conserved - Ser, Leu, Tyr
- alpha-helix H82
Asn69 - not conserved - Asn, Ala, Glu
- alpha-helix H82
Gly70 - not conserved - Gly, Asp, Thr
Asp71 - not conserved - Pro, Asn, Thr
- non-reacting group EAC
Thr72 - not conserved - Ala, Asn, Leu
- alpha-helix H83
Ser73 - not conserved - Ser, Ile, Phe
- alpha-helix H83
Leu74 - not conserved - Glu, Asn, Gly
- alpha-helix H83
Leu75 - not conserved - Thr, Asn
- alpha-helix H83
Ser76 - not conserved - Leu, Thr, Lys
- alpha-helix H83
Thr77 - conserved - Leu, Thr
- alpha-helix H83
Ile78 - not conserved - Ile, Gln, Val
- alpha-helix H83
Glu79 - conserved - Leu, Ile, Glu
- non-reacting group or not protected by acarbose EAC
- alpha-helix H83
Asn80 - conserved
- alpha-helix H83
Tyr81 - conserved - Val, Thr, Tyr
- non-reacting group EAC
- alpha-helix H83
Ile82 - conserved - Ser, Ile
- alpha-helix H83
Ser83 - not conserved - Phe, Tyr, Leu
- alpha-helix H83
Ala84 - not conserved - His, Asn, Lys
- alpha-helix H83
Gln85 - conserved - Leu, Gln, Asp
- alpha-helix H83
Ala86 - conserved - Glu, Ala, Tyr
- alpha-helix H83
Ile87 - conserved - Arg, Val, Ile
- alpha-helix H83
Val88 - conserved - Thr, Ser, Val
- alpha-helix H83
Gln89 - not conserved - Asn, Ser
- alpha-helix H83
Gly90 - conserved - Asn, Phe, Gly
- alpha-helix H83
Ile91 - not conserved - Thr, Pro, Ser
Ser92 - not conserved - Ser, Leu, Val
Asn93 - not conserved - Ala, Ser, Lys
Pro94 - not conserved - Gly, Phe, Thr
Ser95 - not conserved - Ile, Gln
Gly96 - conserved - Asp, Ser
Asp97 - not conserved - Asp, Tyr, Thr
- non-reacting group EAC
Leu98 - not conserved - Thr, Glu, Ser
Ser99 - conserved - Asn, Ser, Thr
Ser100 - not conserved - Asp, Val
Gly101 - not conserved - Thr, His, Cys
Ala102 - not conserved - Val, Lys, Asn
Gly103 - not conserved - Gly, Ala, Cys
Region S2
Leu104 - conserved
Gly105 - conserved
Glu106 - conserved - Asp
- not protected by acarbose EAC
Pro107 - conserved
Lys108 - conserved
- near Trp120
Phe109 - conserved - Trp
Asn110 - conserved
- far away - subsite 5 ??
Val111 - not conserved - Thr, Pro, Val
Asp112 - conserved
- non-reactive or in part protected by acarbose EAC
Glu113 - not conserved - Gly , Asn
- non-reacting group EAC
Thr114 - conserved - Ser
Ala115 - conserved - Gly
- conserved in alpha-amylases
Tyr116 - conserved - Phe
- conserved in alpha-amylases
- partial oxydation with N-bromosuccimidine when unprotected
- near Trp120
*Ala116 - decrease 2 to 7x of ko
- Km (moderate decrease for G2 + iG2, 3x for G7)
- decrease 2x selectivity of G7 over G2)
Thr117 - conserved
- conserved in alpha-amylases
Gly118 - conserved - Glu
- conserved in alpha-amylases
Ser119 - not conserved - Ala, Pro, Asp
- far away - subsite ??
*Tyr119 - slightly higher ko
- slightly smaller Km
- no selectivity data available yet
Trp120 - conserved
- protected by acarbose N-bromosuccinimide
- essential for activity
- located near subsite 4
- influences subsites 1 and 2
- H-bond Glu179 (w/ Trp52 , Gln124)
- near Trp50, Lys108, Trp116
*His120 - decrease 60 to 220x of ko
- Km (2x decrease for G2 , 1.5X increase for G7)
- increase 1.5x selectivity of G7 over G2
*Leu120 - decrease 46 to 56x of ko
- Km (3x decrease for G2)
- decrease 3x selectivity of G7 over G2
...Sf... *Leu120 - no activity at 30deg. or 50deg. C
*Phe120 - decrease 53 to 81x of ko
- Km (2x decrease for G2)
- small decrease of selectivity of G7 over G2
*Tyr120 - decrease 83 to 191x of ko
- Km (3x decrease for G2, 4x decrease for G7, 11x
decrease for iG2)
- small decrease of selectivity of G7 over G2
- decrease 6x selectivity of iG2 over G2
Gly121 - conserved
Arg122 - conserved
Pro123 - conserved
Gln124 - conserved
- H-bond Glu179 (w/ Trp52 , Trp120)
- H-bond Arg54
Arg125 - conserved - Asn, Arg
- H-bond Arg54 ??
Asp126 - conserved
- non-reacting group EAC
- H-bond Arg54 ??
- H-bond amide-N Gly126
Gly127 - conserved
Pro128 - conserved
- interaction Tyr165
Ala129 - conserved
Leu130 - conserved - Glu
- alpha-helix H84
Arg131 - conserved
- alpha-helix H84
Ala132 - conserved - Ser
- alpha-helix H84
Thr133 - not conserved - Thr, Ile, Tyr
- alpha-helix H84
Ala134 - conserved - Thr
- alpha-helix H84
Region Inter S2 - S3
Met135 - not conserved - Lys, Arg, Phe
- alpha-helix H84
Ile136 - conserved - Tyr
- alpha-helix H84
Gly137 - conserved - Ile, Leu, Gly
- alpha-helix H84
Phe138 - not conserved - Phe, Tyr, Glu
- alpha-helix H84
Gly139 - not conserved - Ile, Gly, Ala
- alpha-helix H84
Gln140 - not conserved - Lys, Gln, Asp
- alpha-helix H84
Trp141 - not conserved - Gln, Leu, Ser
- non-reacting group N-bromosuccinimide
- alpha-helix H84
Leu142 - not conserved - Ser, Tyr, Val
- alpha-helix H84
Leu143 - conserved - Gly
- alpha-helix H84
Asp144 - conserved - Thr, Asp
- alpha-helix H84
Asn145 - conserved - Asp, Asn, Gln
- alpha-helix H84
Gly146 - not conserved - Leu, Thr, Ser
Tyr147 - conserved - Gly, Tyr, Lys
- partial oxydation with N-bromosuccimidine when unprotected
Thr148 - not conserved - Asp, Ala
Ser149 - not conserved - Lys, Ile, Ala
- alpha-helix H85
Thr150 - not conserved - Tyr, Ser
- alpha-helix H85
Ala151 - not conserved - Pro, Tyr, Asn
- alpha-helix H85
Thr152 - not conserved - Thr, Gln, Ser
- alpha-helix H85
Asp153 - conserved - Ser, Asp, Gly
- in part protected by acarbose EAC
- alpha-helix H85
Ile154 - conserved - Thr, Ile
- alpha-helix H85
Val155 - not conserved - Ala, Leu, Glu
- alpha-helix H85
Trp156 - conserved - Asp, Trp, Lys
-non-reacting group N-bromosuccinimide
- alpha-helix H85
Pro157 - conserved - Ile, Pro
- alpha-helix H85
Leu158 - not conserved - Phe, Tyr, Ala
- alpha-helix H85
Val159 - not conserved - Glu, Ile, Lys
- alpha-helix H85
Arg160 - conserved - Phe, Lys
- alpha-helix H85
Asn161 - not conserved - Trp, Pro,Lys
- alpha-helix H85
Region S3
Asp162 - conserved
- non-reacting group EAC
- alpha-helix H85
Leu163 - conserved
- alpha-helix H85
Ser164 - not conserved - Arg, Glu , Asp
- alpha-helix H85
Tyr165 - conserved - Phe
- alpha-helix H85
- interaction Pro128
Val166 - conserved - Ile
- alpha-helix H85
Ala167 - conserved - Ile, Val, Ala
- alpha-helix H85
Gln168 - not conserved - Asp, Gly, Asn
- alpha-helix H85
Tyr169 - not conserved - Tyr, His, Val
- partial oxydation with N-bromosuccimidine when unprotected
Trp170 - conserved
- protected only with isomaltose N-bromosuccinimide
- H-bond Gln216
Asn171 - conserved - Ser, Asp
- N-glycosylation site
Gln172 - not conserved - Ser, Asn
Thr173 - not conserved - Thr, Ser, Gly
- essential for N-glycolysation
Gly174 - conserved - Cys
Tyr175 - conserved - Phe, Tyr
- partial oxydation with N-bromosuccimidine when unprotected
Asp176 - conserved
- protected by acarbose EAC
- catalytic base of pKa 2.7 located between subsites 1 and 2
- H-bond amide-N Glu180 ( important conformational role)
- H2O-bridges Ser185 ?? , Asp247 ??
- near Trp52 (Arg54, ??)
* Glu176 - activity observed
* Asn176 - decrease 10 to 20x in ko
- Km - increase 3 to 4x
- decreases 2x selectivity toward G2 over iG2
- increases 2x selectivity toward G6 over G2
aa176.5[*] - insertion based on homology with alpha-amylases
* Gly176.5 - no activity observed
* Asp176.5 - not studied yet
Leu177 - conserved
- affects binding to subsites 1 and 2 (+ Trp178 and Asn182)
- also expected to be involved in the Trp120 conformation
- fills space in cleft
* His177 - decrease ko (10x iG2 , 6x G2 + G6)
- increase KM ( 1.5x G6 + iG2 , 3x G2)
- no change in selectivity for G2/iG2
- increases 2x selectivity toward G6 over G2
* Asp177 - activity observed
- hydrophobic ring increases selectivity for G2 hydrolysis ??
* Phe177 and Trp177 planned
Trp178 - conserved
- not protected by acarbose N-bromosuccinimide
- bond with Arg241 (with Tyr306)
- H-bond with Asp245
- stacked with Arg305 (which is stacked with Trp317)
* Arg178 - based on a-amylase homology
- decrease 4 to 8x for ko
- Km (slight decrease G2 and increase G6 , 2x iG6)
- increased 2x selectivity toward G2 over iG2
- decreased 2x selectivity toward G6 over G2
* Asp178 - no activity observed
* His178 , Phe178 and Tyr178 planned (bulky side-chains)
Glu179 - conserved
- protected by acarbose EAC
- general acid catalyst with pKa 5.9
- H-bond Trp52
- H-bond Trp120
- H-bond Gln124
* Gln179 - decrease of ko 20x for G7
- Km (slight decrease for G7)
- no selectivity data
* Asp179 - no activity observed
Glu180 - conserved
- protected by acarbose EAC
- envolved in substrate binding -interacts with 2'- and 3'-OH of G2
- H-bond Arg305
- H-bond Tyr306
* Gln180 - decrease 2 to 10x for ko
- increase 3 to 4x for Km
- increased 2x selectivity toward G7 over G2
- decrease 30x selectivity toward G2 over iG2
* Asp180 - activity observed
Val181 - conserved - Asn
* Asp181 - activity observed
Asn182 - conserved - Gln
- H-bond carbonyl-O Gly174
* Ala182 - based on homology with a-amylase
- slight decrease for ko
??? - Km (slight decrease for G2, slight increase for G7,
2x increase for iG2)
- selectivity increase of G2 over iG2
* Asp182 - not studied yet
Gly183 - conserved
* Lys183 - slightly higher ko
- slightly smaller Km
- no selectivity data available yet
Ser184 - not conserved - Met, Val, Arg
- alpha-helix H86
?? * His184 - no kinetic data available
Ser185 - conserved - His, Ser
- alpha-helix H86
- H-bond Thr188
Phe186 - conserved
- near bottom of cleft
- alpha-helix H86
Phe187 - conserved - Tyr
- near bottom of cleft
- alpha-helix H86
Thr188 - conserved
- alpha-helix H86
- H-bond amide-N Gly174 ??
- H-bond Ser185
Ile189 - conserved - Leu, Ser, Ile
- alpha-helix H86
Ala190 - conserved - Leu, Met, Ala
- alpha-helix H86
Val191 - conserved
- alpha-helix H86
Gln192 - conserved - Met
- alpha-helix H86
His193 - not conserved - Leu, Gln, Arg
- non-reacting group EAC
- alpha-helix H86
Arg194 - not conserved - Lys, Ser
- alpha-helix H86
Ala195 - conserved - Gly
- alpha-helix H86
Leu196 - conserved - Val
- alpha-helix H86
Region Inter S3 - S4
Val197 - not conserved - Asp, Ala, Leu
- alpha-helix H86
Glu198 - not conserved - Tyr, Arg, Lys, Leu
- non-reacting group EAC
- alpha-helix H86
Gly199 - not conserved - Gly, Ser, Ala
- alpha-helix H86
Ser200 - not conserved - Leu, Val, Ala
- alpha-helix H86
Ala201 - not conserved - Asp, Ser
- alpha-helix H86
Phe202 - not conserved - Phe, Tyr, Ile
- alpha-helix H86
Ala203 - conserved - Phe
- alpha-helix H86
Thr204 - not conserved - Lys, Asn
- alpha-helix H86
Ala205 - conserved - Ser, Arg
- alpha-helix H86
Val206 - not conserved - Ser, Phe, Asn
- alpha-helix H86
Gly207 - not conserved - Gly, Glu, Asp
Ser208 - not conserved - Asp, Arg
Ser209 - conserved - Gly
Cys210 - not conserved - Ser, Asp, Thr
- S-bond with Cys213
- alpha-helix H87
Ser211 - not conserved - Pro, Phe, Arg
- alpha-helix H87
Trp212 - not conserved - Ala, Phe
- protected by acarbose, maltose and isomaltose N-bromosuccinimide
- alpha-helix H87
Cys213 - not conserved - Val, Asn, Ser
- S-bond with Cys210
- alpha-helix H87
Asp214 - not conserved - Thr, Glu
- non-reacting group EAC
- alpha-helix H87
Ser215 - conserved - Arg
- alpha-helix H87
Gln216 - conserved - Thr
- alpha-helix H87
- H-bond Trp170
Ala217 - conserved - Thr
- alpha-helix H87
Pro218 - not conserved - Ser, Arg
- alpha-helix H87
Glu219 - not conserved - Thr, Arg
- non-reacting group EAC
- alpha-helix H87
Ile220 - not conserved - Ile, Asp, Leu
- alpha-helix H87
Leu221 - not conserved - Ile, Glu, Ala
- alpha-helix H87
Cys222 - conserved - Ser, Asn, Cys
- S-bond with Cys449
- alpha-helix H87
Tyr223 - conserved - Arg, Tyr
- partial oxydation with N-bromosuccimidine when unprotected
- alpha-helix H87
Leu224 - not conserved - Leu, Phe, Ile
- alpha-helix H87
Gln225 - not conserved - Leu, Ser
- alpha-helix H87
Ser226 - conserved - Val
- alpha-helix H87
Phe227 - not conserved - Phe, Asp, Gly
- alpha-helix H87
Trp228 - not conserved - Trp, Pro, Ser
- non-reacting group
- alpha-helix H87
Thr229 - not conserved - Ala, Asp, Val
Gly230 - not conserved - Gly, Asn, Ser
Ser231 - conserved - Gly, Asp
Phe232 - conserved - Asn
Ile233 - conserved - Val, Asn
Leu234 - conserved - Asn, Leu, Trp
Ala235 - not conserved - Gly, Thr, Ile
Asn236 - not conserved - Arg, Asp, Gln
Phe237 - not conserved - Val, Tyr
Asp238 - conserved - Asn, Asp, Ser
- non-reacting group EAC
Ser239 - not conserved - Tyr, His, Gln
Ser240 - conserverd - Ile, Ser, Arg
Arg241 - conserved - Pro, Arg
- H-bond Glu180
- H-bond Tyr306
Ser242 - not conserved - Glu, Gly, Thr
- H-bond Glu308
Gly243 - not conserved - Gly, Thr, Asn
Lys244 - conserved - Pro, Gly, Lys
Asp245 - not conserved - Ser, Leu
- non-reacting group EAC
- in vicinity to be catalytic
- H-bonds to Trp178
Ala246 - conserved - Arg, Lys, Ala, Thr
- alpha-helix H8
Asn247 - not conserved - Ser, Gln, Lys
- alpha-helix H8
- close (< 5 Å) Asp176
Thr248 - conserved - Gly, Thr
- alpha-helix H8
Leu249 - conserved
- alpha-helix H8
Leu250 - conserved - Asp, Leu
- alpha-helix H8
Gly251 - not conserved - Ile, Ser,Val
- alpha-helix H8
Ser252 - conserved - Ala
- alpha-helix H8
Ile253 - conserved - Thr, Ile
- alpha-helix H8
His254 - conserved - Leu, Tyr, His
- iodoacetamide-modified
- behind Trp417 stabilization ?
- directing binding ?
- bottom of cleft ?
- alpha-helix H8
Thr255 - conserved - Leu, Thr, Ile
- alpha-helix H8
Phe256 - conserved - Ala, Phe, Gly
- alpha-helix H8
Asp257 - conserved - Ala, Asp, Pro
- non-reacting group EAC
Pro258 - conserved - Asn, Pro, Leu
Glu259 - conserved - His, Glu, Leu
- in part protected by acarbose EAC
Ala260 - conserved - Asp, Ala, Gly
Ala261 - conserved - Ile, Ser
Cys262 - not conserved - Pro, Gly, Val
- S-bond with Cys270
Asp263 - not conserved - Asp, Ser, Glu
- non-reacting group EAC
Asp264 - not conserved - Asp, Ala, Ser
- non-reacting group EAC
Ser265 - conserved - Gly
Thr266 - not conserved - His, Ser, Phe
- H-bond Glu439
Phe267 - not conserved - Phe, Leu, Thr
Gln268 - conserved - Pro, Thr, Gln
Pro269 - conserved - Phe, Pro
Cys270 - conserved - Asp, Cys, Gly
- S-bond with Cys262
Ser271 - conserved - Ser, Ile, Val
Pro272 - not conserved - Asn, Asp, Glu
- alpha-helix H9
Arg273 - not conserved - Asp, Asn, Lys
- alpha-helix H9
Ala274 - not conserved - Pro, Glu, Ile
- alpha-helix H9
Leu275 - not conserved - Leu, Ala, Tyr
- alpha-helix H9
Ala276 - conserved - Val, Ala
Asn277 - conserved - Leu, Asn, Thr
- alpha-helix H9
His278 - not conserved - Asn, Gln, Ala
- not protected by acarbose EAC
- alpha-helix H9
Lys279 - not conserved - Thr, Val, Ser
- alpha-helix H9
Glu280 - not conserved - Leu, Ala, Tyr
- non-reacting or not protected by acarbose EAC
- alpha-helix H9
Val281 - not conserved - Val, His, Tyr
- alpha-helix H9
Val282 - not conserved - His, Leu, Glu
- alpha-helix H9
Asp283 - conserved - Leu, Asp
- non-reacting group EAC
- alpha-helix H9
Ser284 - not conserved - Ser, Met, Leu
- alpha-helix H9
Phe285 - not conserved - Phe, Leu, Glu
- alpha-helix H9
Arg286 - conserved - Ala, Asp
- alpha-helix H9
- H-bond carbonyl-O Asp297 ??
- H-bond carbonyl-O Glu299 ??
Ser287 - conserved - Asn
- alpha-helix H9
Ile288 - conserved - Lys, Leu
- alpha-helix H9
Tyr289 - conserved - Asp
- non-reacting group EAC
- alpha-helix H9
Thr290 - conserved - Pro, Thr, Arg
- alpha-helix H9 ??
Leu291 - conserved - Ile, Leu, Tyr
- alpha-helix H9 ??
Asn292 - conserved - Ser
- alpha-helix H9 ??
Asp293 - conserved - Val, Arg
- non-reacting group EAC
- alpha-helix H9 ??
Gly294 - not conserved - Asn, Ser
- alpha-helix H9 ??
Leu295 - conserved - Ser, Leu
- alpha-helix H9 ??
Ser296 -not conserved - Lys, Ala, Pro
Asp297 - not conserved - Asn, Tyr, Ser
- non-reacting group EAC
Ser298 - not conserved - Ser, Ala, Tyr
Glu299 - not conserved - Thr, Ala, Leu
- non-reacting group EAC
Region S4
Ala300 - conserved - Gly, Ala
Val301 - not conserved - Ile, Ala, Asn
Ala302 - conserved - Ser
Val303 - not conserved - Ile, Leu
Gly304 - conserved
- rigid point
Arg305 - conserved
- 2 H-bonds Asp309
- H-bond Glu180
Tyr306 - conserved
- non-reacting-group EAC
- H-bond Glu180
- H-bond Arg241
* Phe306 - decrease 1.5x ko
- increase 2x Km
- no selectivity data yet
Pro307 - conserved
Glu308 - conserved
- non-reacting group EAC
- H-bond Ser242
Asp309 - conserved
- non-reacting group EAC
- 2 H-bonds Asp305
- H-bond Tyr311
* Asn309 - decrease 20x ko
- increase 4x Km
- no selectivity data yet
Thr310 - conserved - Val, Thr
Tyr311 - conserved
- non-reacting group EAC
- H-bond Tyr48
- H-bond Asp309
- cleft
Tyr312 - not conserved - Asn, Asp
- non-reacting group EAC
- cleft
Asn313 - not conserved - Asn, Tyr, Asp
Gly314 - conserved
Asn315 - conserved
- H-bond with Gln401 (structure stabilization ?)
Pro316 - conserved - Ser
Trp317 - conserved
- non-reacting group N-bromosuccinimide
- stacked with Arg305 (which is stacked with Trp178)
- stacked with Glu400 ??
Phe318 - conserved - Val
- alpha-helix H10
Leu319 - conserved
- alpha-helix H10
Cys320 - conserved - Ala , Cys
- alpha-helix H10
Region Inter S4 - S5
Thr321 - conserved - Val
- alpha-helix H10
Leu322 - not conserved - Thr, Ala, Tyr
- alpha-helix H10
Ala323 - conserved - Gly
- alpha-helix H10
Ala324 - not conserved - Ala, Ser, Tyr
- alpha-helix H10
Ala325 - not conserved - Ala, Thr, Gln
- alpha-helix H10
Glu326 - not conserved - Glu, Thr, Val
- alpha-helix H10
Gln327 - conserved - Lys
- alpha-helix H10
Leu328 - conserved
- alpha-helix H10
Tyr329 - conserved
- partial oxidation with N-bromosuccinimide when unprotected
- alpha-helix H10
Asp330 - conserved - Arg, Asp
- not protected by acarbose EAC
- alpha-helix H10
Ala331 - conserved - His
- alpha-helix H10
Leu332 - conserved - Ile, Lys, Leu
- alpha-helix H10
Tyr333 - conserved - Ser, Arg, Tyr
- non-reacting group EAC
- alpha-helix H10
Gln334 - conserved - Ser
- alpha-helix H10
Trp335 - not conserved - Trp, His, Asn
- non-reacting group N-bromosuccinimide
- alpha-helix H10
Asp336 - conserved - Ile
- non-reacting group EAC
- alpha-helix H10
Lys337 - not conserved - Leu, Ile, Gln
- alpha-helix H10
Gln338 - not conserved - Val, Asn
- alpha-helix H10
Gly339 - not conserved - Gly, Val
- beta-strand 1
Ser340 - not conserved - Pro, Thr, Gly
- beta-strand 1
Leu341 - not conserved - Met, Val, Ile
- beta-strand 1
Glu342 - conserved - Asn, Thr, Glu
- non-reacting group EAC
- beta-strand 1
Val343 - not conserved - Val, Asn, Lys
- beta-strand 1
Thr344 - not conserved - Asp, Ser
Asp345 - not conserved - Cys, Ser
- non-reacting group EAC
Val346 - not conserved - Ile, Ser
- alpha-helix H11
Ser347 - conserved - Asn, Ser
- alpha-helix H11
Leu348 - not conserved - Leu, Ala, Tyr
- alpha-helix H11
Asp349 - not conserved - Asp, Phe, Pro
- non-reacting group EAC
- alpha-helix H11
Phe350 - conserved - Trp
- alpha-helix H11
Phe351 - conserved - Ser
- alpha-helix H11
Lys352 - not conserved - Lys, Glu, Asn
- alpha-helix H11
Ala353 - not conserved - Lys, Leu
- alpha-helix H11
Leu354 - not conserved - Val, Tyr, Phe
- alpha-helix H11
Tyr355 - not conserved - Phe, Ile, Asp
- partial oxidation with N-bromosuccinimide when unprotected
- non-reacting group EAC
Ser356 - conserved - Val
Asp357 - not conserved - Asp, Asn, Ser
- non-reacting group EAC
Ala358 - conserved - Leu, Ala
Ala359 - conserved - Thr, Ser, Ala
Thr360 - conserved - Ser
Gly361 - not conserved - Gly, Leu, Ile
- beta-strand 1
Thr362 - not conserved - Thr, Asn, Lys
- beta-strand 1
Tyr363 - not conserved - Tyr, Asp, Ile
- partial oxidation with N-bromosuccinimide when unprotected
- non-reacting group EAC
- beta-strand 1
- H-bond carbonyl-O Ala359
Ser364 - not conserved - Glu, Lys, Thr
- beta-strand 1
Ser365 - not conserved - Ser, Gly, Val
- beta-strand 1
Ser366 - not conserved - Gly, Tyr
Ser367 - not conserved - Ser, Leu, Thr
Ser368 - not conserved - Ser, Ile
- alpha-helix H12
Thr369 - not conserved - Asp, Leu
- alpha-helix H12
Tyr370 - conserved - Glu, Tyr, Phe
- non-reacting group EAC
- alpha-helix H12
Ser 371 - conserved - Phe, Ser, Asn
- alpha-helix H12
Ser372 - conserved - Asn, Ser
- alpha-helix H12
Ile373 - conserved - Thr, Leu, Ile
- alpha-helix H12
Val374 - not conserved - Val, Pro, Ala
- alpha-helix H12
Asp375 - conserved - Ala, Gln, Asp
- not protected by acarbose EAC
- alpha-helix H12
Ala376 - not conserved - Phe, Asp, Asn, Ala
- alpha-helix H12
Val377 - conserved - Asn, Ile, Val
- alpha-helix H12
Lys378 - not conserved - Gln, Leu, Ala
- alpha-helix H12
Thr379 - conserved - Leu
- alpha-helix H12
Phe380 - not conserved - Phe, Ile, Ala
- alpha-helix H12
Ala381 - not conserved - Ala, Gln, Gly
- alpha-helix H12
Asp382 - conserved - Lys
- non-reacting group EAC
- alpha-helix H12
Gly383 - not conserved - Ile, Ser, Arg
- alpha-helix H12
Phe384 - conserved
- alpha-helix H12
Val385 - not conserved - Gln, Leu
- alpha-helix H12
Ser386 - conserved - Leu, Ser
- alpha-helix H12
Ile387 - not conserved - Ala, Gln, Thr
- alpha-helix H12
Val388 - conserved - Asp
- alpha-helix H12
Glu389 - not conserved - Ser, Gln, Ile
- in part protected by acarbose EAC
- alpha-helix H12
Thr390 - not conserved - Leu, Phe
- alpha-helix H12
His391 - conserved - Leu
- non-reacting group EAC
- alpha-helix H12
Ala392 - not conserved - Ala, Val, Ile
- alpha-helix H12
Ala393 - not conserved - Lys, Asn, His
Ser394 - not conserved - Leu, Asp, Asn
Asn395[ ] - not conserved - Asn, Lys[*], Asp
- N-glycosylation site
- Carbohydrate H-bond Asp47
- Carbohydrate H-bond Arg413
Region S5
Gly396 - conserved - Ala[*]
Ser397 - conserved - Thr
- essential for N-glycosylation
Met398 - not conserved - Leu, Trp[*]
Ser399 - not conserved - Asn, Ala
Glu400 - conserved
- not protected by acarbose
- lining cleft
- H-bond with Gln401
- H-bond with Tyr48, Ser411 ??
- stacked with Trp317 ??
Gln401 - conserved - Glu
- lining cleft
- H-bond with Glu400
- H-bond with Asn315 (structure stabilization ?)
Tyr402 - not conserved - Thr[*], Leu, Phe
- partial oxydation with N-bromosuccimidine when unprotected
Asp403 - not conserved - Asp, Gly[*], Asn
- not protected by acarbose EAC
Lys404 - not conserved - Arg, Asn[*
]- H-bond Leu291
Ser405 - not conserved - Tyr, Thr
Asp406 - not conserved - Thr
- non-reacting group EAC
Gly407 - conserved
Glu408 - not conserved - Tyr, Leu
- non-reacting group EAC
Gln409 - not conserved - Ser
Leu410 - not conserved - Thr
Ser411 - not conserved - Gly
- lining cleft
- H-bond with Glu400 ??
Ala412 - conserved
Arg413 - conserved - Tyr
- H-bond Carbohydrate Asn395
Asp414 - conserved - Ser
- non-reacting group EAC
- alpha-helix H13
Leu415 - conserved
- alpha-helix H13
Thr416 - conserved
- interaction with Asp55 on free side chain In active site ??
- alpha-helix H13
Trp417 - conserved
- protected by maltose, acarbose and isomaltose
?? - bottom of active site - block action
- stabilized by His254 ?
- alpha-helix H13
Ser418 - conserved
- alpha-helix H13
Tyr419 -not conserved - Ser, His
- partial oxydation with N-bromosuccimidine when unprotected
- alpha-helix H13
Ala420 - conserved - Gly
- alpha-helix H13
Ala421 - conserved - Ser
- alpha-helix H13
Leu422 - conserved
- alpha-helix H13
Leu423 - conserved - Ile
- alpha-helix H13
Thr424 - conserved - Glu
- alpha-helix H13
Ala425 - conserved
- alpha-helix H13
Region Pos-S5
Asn426 - not conserved - Ile, Ser
- alpha-helix H13
Asn427 - not conserved - Lys, Tyr
- alpha-helix H13
Arg428 - not conserved - Ala, Leu
- alpha-helix H13
Arg429 - conserved - Lys
- alpha-helix H13
Asn430 - conserved - Ala
- alpha-helix H13
Ser431 - not conserved - Lys, Gly
Val432 - conserved - Gly
Val433 - not conserved - Lys, Pro
Pro434 - conserved - Ala, Pro
Ala435 - conserved - Leu
Ser436 - not conserved - Ala
Trp437 - not matched
- protected by maltose, acarbose and isomaltose N-bromosuccinimide
Gly438 - not matched
Glu439 - not matched
- non-reacting or not protected by acarbose EAC
- alpha-helix N
- H-bond Thr266
Thr440 - not matched
- alpha-helix N
Ser441 - not matched
- O-glycosylation site
- alpha-helix N
Ala442 - not matched
- alpha-helix N
* Thr442 - thermosensitive
- O-glycosylated ??
Ser443 - not matched
- O-glycosylation site
- alpha-helix N
Ser444 - not matched
- O-glycosylation site
- alpha-helix N
Val445 - not matched
- alpha-helix N
Pro446 - not matched
Gly447 - not matched
Thr448 - not matched
Cys449 - not matched
- S-bond with Cys222
Ala450 - not matched
Ala451 - not matched
Thr452 - not matched
- O-glycosylation site
Ser453 - not matched
- O-glycosylation site
Ala454 - not matched
Ile455 - not matched
Gly456 - not matched
Thr457 - not matched
Tyr458 - not matched
- non-reacting group EAC
Ser459 - not matched
- O-glycosylation site
Ser460 - not matched
- O-glycosylation site
Val461 - not matched
Thr462 - not matched
- O-glycosylation site
Val463 - not matched
Thr464 - not matched
- O-glycosylation site
Ser465 - not matched
Trp466 - not matched
- not protected by acarbose N-bromosuccinimide
Pro467 - not matched
Ser468 - not matched
- O-glycosylation site
Ile469 - not matched
Val470 - not matched
Last Changed: 05/20/94