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. Author manuscript; available in PMC: 2012 Nov 8.
Published in final edited form as: Biochemistry. 2011 Oct 11;50(44):9488–9499. doi: 10.1021/bi2009807

Table 1.

Catalytic properties (at 25 ºC, pH 7.5) of MMP-12 variants found to have consequential mutations.

MMP-12 varianta Location
of
mutation		 General Substrate
FS-6
 fEln-100
 Relative
activity of
fEln-100
FS-6
normalized
to
kcat/Km,/M/s kcat,
s−1 		Km,
µM		 kcat/Km
/M/s		 kcat,10
2s−1 		Km
µM		 wtMMP12c:
selectivity
for fEln-
100
wt MMP-12 b 133,800 ± 6000 17.3 ± 0.8 129 ± 4 10690 ± 430 1.06 ± 0.04 1.06 ± 0.04 100.0%
wt MMP-3 b 20,660 ± 620 3.2 ± 0.1 154 ± 5 830 ± 40 0.08 ± 0.01 0.94 ± 0.04 50.3%
M103F N-terminus 146,300 ± 4400 18.7 ± 0.6 128 ± 4 8040 ± 320 0.92 ± 0.04 1.15 ± 0.04 68.8%
R117S Stand I, next to II–III loop 120,800 ± 3600 13.7 ± 0.4 113 ± 3 7130 ± 290 0.72 ± 0.03 1.15 ± 0.04 73.8%
D124Q I-A loop, Ca2+ ligand 136,700 ± 4100 17.1 ± 0.5 125 ± 4 7060 ± 280 1.24 ± 0.05 1.75 ± 0.07 64.6%
M156E II–III loop 135,200 ± 4100 16.9 ± 0.5 125 ± 4 4890 ± 200 1.17 ± 0.05 2.39 ± 0.10 45.3%
I180Sb Bulge-edge in III–IV loop 141,800 ± 4300 24.1 ± 2.2 170 ± 15 2860 ± 140 0.37 ± 0.06 1.3 ± 0.1 25.2%
A182G Strand IV 120,100 ± 3600 15.6 ± 0.5 130 ± 4 4970 ± 200 0.57 ± 0.02 1.14 ± 0.05 51.8%
D124Q/M156E 134,700 ± 4000 16.7 ± 0.5 124 ± 4 4210 ± 200 1.43 ± 0.06 3.40 ± 0.14 39.1%
D124Q/I180S 67,230 ± 2020 16.2 ± 0.5 241 ± 7 1855 ± 80 0.24 ± 0.01 1.27 ± 0.05 34.5%
D124Q/A182G 54,610 ± 1640 10.6 ± 0.3 194 ± 6 1660 ± 70 0.18 ± 0.01 1.09 ± 0.04 38.1%
M156E/I180S 57,420 ± 1720 6.8 ± 0.2 119 ± 4 1310 ± 50 0.52 ± 0.02 3.99 ± 0.16 28.6%
145,400 ± 4400 17.2 ± 0.5 118 ± 4 4650 ± 190 2.16 ± 0.09 4.66 ± 0.19 40.0%
D124Q/M156E/I180S 53,660 ± 1600 9.7 ± 0.3 180 ± 5 829 ± 33 0.10 ± 0.01 1.19 ± 0.05 19.3%
D124Q/M156E/A182G 41,570 ± 1250 8.4 ± 0.3 202 ± 6 593 ± 24 0.10 ± 0.01 1.71 ± 0.07 17.9%
D124Q/M156E/F185Y 133,400 ± 4000 18 ± 0.5 135 ± 4 3570 ± 140 0.48 ± 0.02 1.35 ± 0.05 33.5%
D124Q/M156E/T205K 159,700 ± 4800 18 ± 0.5 113 ± 4 4500 ± 180 0.65 ± 0.03 1.45 ± 0.06 35.3%
a

Each mutation is found at the equivalent position in a less elastolytic homologue such as MMP-3, −8, −10, −11, −27, or MT-MMP.

b

previously reported in ref (16).

c

heights from Fig. 2a of black columns divided by hatched columns, i.e. kcat/Km,fEln100/ kcat/Kmfs6 after normalizing each kcat/Km to the wild type