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. Author manuscript; available in PMC: 2019 Feb 19.

Published in final edited form as: Biochemistry. 2019 Jan 29;58(7):987–996. doi: 10.1021/acs.biochem.8b01246

Table 1.

Attributes of RNase 1 Glycoforms

			m/z (Da)			k_cat/K_M^c
Source	RNase 1^a	Glycans	expected	observed	T_m (°C)^b	10⁶ M^–1s^–1	%
E. coli	NNN	0	14705.52^d	14698.35	55.9 ± 0.2	1.9 ± 0.2	100
E. coli	QQQ	0	14747.60^d	14740.03	56.6 ± 0.3	1.9 ± 0.1	100
P. pastoris	QQQ	0	14616.41	14608.88	56.4 ± 0.1	1.9 ± 0.1	100
P. pastoris	NQQ	1	15818.38	15811.70	58.6 ± 0.3	1.5 ± 0.1	79
P. pastoris	QNQ	1	15818.38	15811.91	56.8 ± 0.1	0.74 ± 0.07	39
P. pastoris	QQN	1	15818.38	15811.85	57.2 ± 0.2	0.30 ± 0.04	16
P. pastoris	NNQ	2	17020.36	17015.05	59.1 ± 0.2	0.72 ± 0.04	38
P. pastoris	NQN	2	17020.36	17014.49	58.7 ± 0.2	0.63 ± 0.03	33
P. pastoris	QNN	2	17020.36	17015.57	57.3 ± 0.1	0.21 ± 0.03	11
P. pastoris	NNN	3	18222.33	18218.39	59.4 ± 0.3	0.19 ± 0.09	10

^a

Residues at positions 34, 76, and 88 (Figure 1D).

^b

Values were determined by DSF at pH 7.4.

^c

Values were determined by fluorescence spectroscopy for the cleavage of 6-FAM–dArUdAdA–6-TAMRA at pH 7.5.

^d

The proteins produced in E. coli have an N-terminal methionine residue.