Mutagenesis of Lys35, Tyr73, Lys74, Glu80, Asp82, Asn95, Lys130, Lys135, or Lys136, identified by comprehensive Ala-scanning
| Variant . | Spec. Act. (kU/mg) . | Antibody absorption (%) . |
|---|---|---|
| SakSTAR | 120 | 95 |
| SakSTAR(K35A) | 230 | 91 |
| SakSTAR(K35E) | 160 | 95 |
| SakSTAR(K35Q) | 69 | 95 |
| SakSTAR(W66A) | <5 | 85 |
| SakSTAR(Y73A) | <5 | 63 |
| SakSTAR(Y73F) | 31 | 93 |
| SakSTAR(Y73H) | <5 | 76 |
| SakSTAR(Y73L) | <5 | 81 |
| SakSTAR(Y73S) | <5 | 86 |
| SakSTAR(Y73W) | 27 | 73 |
| SakSTAR(K74A) | 69 | 64 |
| SakSTAR(K74E) | <5 | 83 |
| SakSTAR(K74N) | 39 | 63 |
| SakSTAR(K74Q) | 110 | 70 |
| SakSTAR(K74R) | 150 | 75 |
| SakSTAR(E80A,D82A) | 130 | 89 |
| SakSTAR(E80A) | 160 | 94 |
| SakSTAR(D82A) | 160 | 95 |
| SakSTAR(N95A) | 260 | 95 |
| SakSTAR(N95E) | 79 | 95 |
| SakSTAR(N95G) | 160 | 95 |
| SakSTAR(N95K) | 180 | 95 |
| SakSTAR(K130A) | 280 | 92 |
| SakSTAR(K130T) | 290 | 92 |
| SakSTAR(K135A) | 410 | 95 |
| SakSTAR(K135F) | 64 | 95 |
| SakSTAR(K135R) | 230 | 95 |
| SakSTAR(K136A) | 180 | 91 |
| SakSTAR(K136R) | 94 | 95 |
| SakSTAR(K136F) | 88 | 84 |
| SakSTAR(∇137K) | 1600 | 95 |
| SakSTAR(∇137A) | 88 | 95 |
| SakSTAR(K136A,∇137K) | 1800 | 88 |
| Variant . | Spec. Act. (kU/mg) . | Antibody absorption (%) . |
|---|---|---|
| SakSTAR | 120 | 95 |
| SakSTAR(K35A) | 230 | 91 |
| SakSTAR(K35E) | 160 | 95 |
| SakSTAR(K35Q) | 69 | 95 |
| SakSTAR(W66A) | <5 | 85 |
| SakSTAR(Y73A) | <5 | 63 |
| SakSTAR(Y73F) | 31 | 93 |
| SakSTAR(Y73H) | <5 | 76 |
| SakSTAR(Y73L) | <5 | 81 |
| SakSTAR(Y73S) | <5 | 86 |
| SakSTAR(Y73W) | 27 | 73 |
| SakSTAR(K74A) | 69 | 64 |
| SakSTAR(K74E) | <5 | 83 |
| SakSTAR(K74N) | 39 | 63 |
| SakSTAR(K74Q) | 110 | 70 |
| SakSTAR(K74R) | 150 | 75 |
| SakSTAR(E80A,D82A) | 130 | 89 |
| SakSTAR(E80A) | 160 | 94 |
| SakSTAR(D82A) | 160 | 95 |
| SakSTAR(N95A) | 260 | 95 |
| SakSTAR(N95E) | 79 | 95 |
| SakSTAR(N95G) | 160 | 95 |
| SakSTAR(N95K) | 180 | 95 |
| SakSTAR(K130A) | 280 | 92 |
| SakSTAR(K130T) | 290 | 92 |
| SakSTAR(K135A) | 410 | 95 |
| SakSTAR(K135F) | 64 | 95 |
| SakSTAR(K135R) | 230 | 95 |
| SakSTAR(K136A) | 180 | 91 |
| SakSTAR(K136R) | 94 | 95 |
| SakSTAR(K136F) | 88 | 84 |
| SakSTAR(∇137K) | 1600 | 95 |
| SakSTAR(∇137A) | 88 | 95 |
| SakSTAR(K136A,∇137K) | 1800 | 88 |
Specific activity (Spec. Act.) ≥ 100 kU/mg is represented in bold type. Absorption of antibodies (in %) from pooled immunized patient plasma; values ≤ 85% are represented in bold type. Variants of wild-type SakSTAR are represented by the substituted amino acids in single letter symbols, followed by their position number in the mature staphylokinase sequence (136 amino acids) and by the substituting amino acids in single letter symbols. Added amino acids are indicated by ∇, followed by their position number and their identity in single letter symbols.