References
1. Wiskerchen M, Muesing MA (1995) Identification and characterization of a temperaturesensitive mutant of human immunodeficiency
virus type 1 by alanine scanning mutagenesis of
the integrase gene. J Virol 69(1):597–601
2. Williams PF, Mynarcik DC, Yu GQ, Whittaker
J (1995) Mapping of an NH2-terminal ligand
binding site of the insulin receptor by alanine
scanning mutagenesis. J Biol Chem 270
(7):3012–3016
3. Blaber M, Baase WA, Gassner N, Matthews BW
(1995) Alanine scanning mutagenesis of the
alpha-helix 115-123 of phage T4 lysozyme:
effects on structure, stability and the binding
of solvent. J Mol Biol 246(2):317–330
4. Manglik A, Kruse AC, Kobilka TS, Thian FS,
Mathiesen JM, Sunahara RK, Pardo L, Weis
WI, Kobilka BK, Granier S (2012) Crystal
structure of the micro-opioid receptor bound
to a morphinan antagonist. Nature 485
(7398):321–326
5. Rasmussen SG, Choi HJ, Rosenbaum DM,
Kobilka TS, Thian FS, Edwards PC,
Burghammer M, Ratnala VR, Sanishvili R,
Fischetti RF, Schertler GF, Weis WI, Kobilka
BK (2007) Crystal structure of the human
beta2 adrenergic G-protein-coupled receptor.
Nature 450(7168):383–387
6. Cherezov V, Rosenbaum DM, Hanson MA,
Rasmussen SG, Thian FS, Kobilka TS, Choi
HJ, Kuhn P, Weis WI, Kobilka BK, Stevens
RC (2007) High-resolution crystal structure
of an engineered human beta2-adrenergic G
protein-coupled receptor. Science 318
(5854):1258–1265
7. Rasmussen SG, DeVree BT, Zou Y, Kruse AC,
Chung KY, Kobilka TS, Thian FS, Chae PS,
Pardon E, Calinski D, Mathiesen JM, Shah
ST, Lyons JA, Caffrey M, Gellman SH,
Steyaert J, Skiniotis G, Weis WI, Sunahara
RK, Kobilka BK (2011) Crystal structure of
the beta2 adrenergic receptor-Gs protein complex. Nature 477(7366):549–555
8. Kobilka B, Schertler GF (2008) New Gprotein-coupled receptor crystal structures:
insights and limitations. Trends Pharmacol Sci
29(2):79–83
9. Stevenson HP, DePonte DP, Makhov AM,
Conway JF, Zeldin OB, Boutet S, Calero G,
Cohen AE (2014) transmission electron
microscopy as a tool for nanocrystal characterization pre- and post-injector. Philos Trans R
Soc Lond Ser B Biol Sci 369(1647):20130322
10. Stevenson HP, Makhov AM, Calero M,
Edwards AL, Zeldin OB, Mathews II, Lin G,
Barnes CO, Santamaria H, Ross TM, Soltis
SM, Khosla C, Nagarajan V, Conway JF,
Cohen AE, Calero G (2014) Use of transmission electron microscopy to identify nanocrystals of challenging protein targets. Proc Natl
Acad Sci U S A 111(23):8470–8475
11. Barnes CO, Kovaleva EG, Fu X, Stevenson HP,
Brewster AS, DePonte DP, Baxter EL, Cohen
AE, Calero G (2016) Assessment of microcrystal quality by transmission electron microscopy
for efficient serial femtosecond crystallography.
Arch Biochem Biophys 602:61–68
12. Stevenson HP, Lin G, Barnes CO,
Sutkeviciute I, Krzysiak T, Weiss SC,
Reynolds S, Wu Y, Nagarajan V, Makhov AM,
Lawrence R, Lamm E, Clark L, Gardella TJ,
Hogue BG, Ogata CM, Ahn J, Gronenborn
AM, Conway JF, Vilardaga JP, Cohen AE,
Calero G (2016) Transmission electron
microscopy for the evaluation and optimization of crystal growth. Acta Crystallogr D
Struct Biol 72(Pt 5):603–615
13. Heras B, Martin JL (2005) Post-crystallization
treatments for improving diffraction quality of
protein crystals. Acta Crystallogr D Biol Crystallogr 61(Pt 9):1173–1180
14. Newman J (2006) A review of techniques for
maximizing diffraction from a protein crystal in
stilla. Acta Crystallogr D Biol Crystallogr 62
(Pt 1):27–31
15. Russo Krauss I, Sica F, Mattia CA, Merlino A
(2012) Increasing the X-ray diffraction power
of protein crystals by dehydration: the case of
bovine serum albumin and a survey of literature
data. Int J Mol Sci 13(3):3782–3800
16. Lin G, Weiss S, Vergara S, Calero G (2019)
Transcription with a laser: radiation-damagefree diffraction of RNA polymerase II crystals.
Methods 159–160:23
Detection of Microcrystals for CryoEM
307
1. Wiskerchen M, Muesing MA (1995) Identification and characterization of a temperaturesensitive mutant of human immunodeficiency
virus type 1 by alanine scanning mutagenesis of
the integrase gene. J Virol 69(1):597–601
2. Williams PF, Mynarcik DC, Yu GQ, Whittaker
J (1995) Mapping of an NH2-terminal ligand
binding site of the insulin receptor by alanine
scanning mutagenesis. J Biol Chem 270
(7):3012–3016
3. Blaber M, Baase WA, Gassner N, Matthews BW
(1995) Alanine scanning mutagenesis of the
alpha-helix 115-123 of phage T4 lysozyme:
effects on structure, stability and the binding
of solvent. J Mol Biol 246(2):317–330
4. Manglik A, Kruse AC, Kobilka TS, Thian FS,
Mathiesen JM, Sunahara RK, Pardo L, Weis
WI, Kobilka BK, Granier S (2012) Crystal
structure of the micro-opioid receptor bound
to a morphinan antagonist. Nature 485
(7398):321–326
5. Rasmussen SG, Choi HJ, Rosenbaum DM,
Kobilka TS, Thian FS, Edwards PC,
Burghammer M, Ratnala VR, Sanishvili R,
Fischetti RF, Schertler GF, Weis WI, Kobilka
BK (2007) Crystal structure of the human
beta2 adrenergic G-protein-coupled receptor.
Nature 450(7168):383–387
6. Cherezov V, Rosenbaum DM, Hanson MA,
Rasmussen SG, Thian FS, Kobilka TS, Choi
HJ, Kuhn P, Weis WI, Kobilka BK, Stevens
RC (2007) High-resolution crystal structure
of an engineered human beta2-adrenergic G
protein-coupled receptor. Science 318
(5854):1258–1265
7. Rasmussen SG, DeVree BT, Zou Y, Kruse AC,
Chung KY, Kobilka TS, Thian FS, Chae PS,
Pardon E, Calinski D, Mathiesen JM, Shah
ST, Lyons JA, Caffrey M, Gellman SH,
Steyaert J, Skiniotis G, Weis WI, Sunahara
RK, Kobilka BK (2011) Crystal structure of
the beta2 adrenergic receptor-Gs protein complex. Nature 477(7366):549–555
8. Kobilka B, Schertler GF (2008) New Gprotein-coupled receptor crystal structures:
insights and limitations. Trends Pharmacol Sci
29(2):79–83
9. Stevenson HP, DePonte DP, Makhov AM,
Conway JF, Zeldin OB, Boutet S, Calero G,
Cohen AE (2014) transmission electron
microscopy as a tool for nanocrystal characterization pre- and post-injector. Philos Trans R
Soc Lond Ser B Biol Sci 369(1647):20130322
10. Stevenson HP, Makhov AM, Calero M,
Edwards AL, Zeldin OB, Mathews II, Lin G,
Barnes CO, Santamaria H, Ross TM, Soltis
SM, Khosla C, Nagarajan V, Conway JF,
Cohen AE, Calero G (2014) Use of transmission electron microscopy to identify nanocrystals of challenging protein targets. Proc Natl
Acad Sci U S A 111(23):8470–8475
11. Barnes CO, Kovaleva EG, Fu X, Stevenson HP,
Brewster AS, DePonte DP, Baxter EL, Cohen
AE, Calero G (2016) Assessment of microcrystal quality by transmission electron microscopy
for efficient serial femtosecond crystallography.
Arch Biochem Biophys 602:61–68
12. Stevenson HP, Lin G, Barnes CO,
Sutkeviciute I, Krzysiak T, Weiss SC,
Reynolds S, Wu Y, Nagarajan V, Makhov AM,
Lawrence R, Lamm E, Clark L, Gardella TJ,
Hogue BG, Ogata CM, Ahn J, Gronenborn
AM, Conway JF, Vilardaga JP, Cohen AE,
Calero G (2016) Transmission electron
microscopy for the evaluation and optimization of crystal growth. Acta Crystallogr D
Struct Biol 72(Pt 5):603–615
13. Heras B, Martin JL (2005) Post-crystallization
treatments for improving diffraction quality of
protein crystals. Acta Crystallogr D Biol Crystallogr 61(Pt 9):1173–1180
14. Newman J (2006) A review of techniques for
maximizing diffraction from a protein crystal in
stilla. Acta Crystallogr D Biol Crystallogr 62
(Pt 1):27–31
15. Russo Krauss I, Sica F, Mattia CA, Merlino A
(2012) Increasing the X-ray diffraction power
of protein crystals by dehydration: the case of
bovine serum albumin and a survey of literature
data. Int J Mol Sci 13(3):3782–3800
16. Lin G, Weiss S, Vergara S, Calero G (2019)
Transcription with a laser: radiation-damagefree diffraction of RNA polymerase II crystals.
Methods 159–160:23
Detection of Microcrystals for CryoEM
307